Insecticidal proteins from plants

ABSTRACT

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

CROSS REFERENCE

This application is a 371 (National Stage) of PCT/US15/14816 filed Feb.6, 2015, which claims the benefit of U.S. Provisional Application No.61/937,288 filed Feb. 7, 2014, which are incorporated herein byreference in their entirety.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

The official copy of the sequence listing is submitted electronicallyvia EFS-Web as an ASCII formatted sequence listing with a file named“20150203_5295 PCT_SeqListing” created on Feb. 3, 2015, and having asize of 1,281 kilobytes and is filed concurrently with thespecification. The sequence listing contained in this ASCII formatteddocument is part of the specification and is herein incorporated byreference in its entirety.

FIELD

This disclosure relates to the field of molecular biology. Provided arenovel genes that encode pesticidal proteins. These pesticidal proteinsand the nucleic acid sequences that encode them are useful in preparingpesticidal formulations and in the production of transgenicpest-resistant plants.

BACKGROUND

Biological control of insect pests of agricultural significance using amicrobial agent, such as fungi, bacteria or another species of insectaffords an environmentally friendly and commercially attractivealternative to synthetic chemical pesticides. Generally speaking, theuse of biopesticides presents a lower risk of pollution andenvironmental hazards and biopesticides provide greater targetspecificity than is characteristic of traditional broad-spectrumchemical insecticides. In addition, biopesticides often cost less toproduce and thus improve economic yield for a wide variety of crops.

Certain species of microorganisms of the genus Bacillus are known topossess pesticidal activity against a range of insect pests includingLepidoptera, Diptera, Coleoptera, Hemiptera and others. Bacillusthuringiensis (Bt) and Bacillus popilliae are among the most successfulbiocontrol agents discovered to date. Insect pathogenicity has also beenattributed to strains of B. larvae, B. lentimorbus, B. sphaericus and B.cereus. Microbial insecticides, particularly those obtained fromBacillus strains, have played an important role in agriculture asalternatives to chemical pest control.

Crop plants have been developed with enhanced insect resistance bygenetically engineering crop plants to produce pesticidal proteins fromBacillus. For example, corn and cotton plants have been geneticallyengineered to produce pesticidal proteins isolated from strains of Bt.These genetically engineered crops are now widely used in agricultureand have provided the farmer with an environmentally friendlyalternative to traditional insect-control methods. While they haveproven to be very successful commercially, these genetically engineered,insect-resistant crop plants provide resistance to only a narrow rangeof the economically important insect pests. In some cases, insects candevelop resistance to different insecticidal compounds, which raises theneed to identify alternative biological control agents for pest control.

Accordingly, there remains a need for new pesticidal proteins withdifferent ranges of insecticidal activity against insect pests, e.g.,insecticidal proteins which are active against a variety of insects inthe order Lepidoptera and the order Coleoptera including but not limitedto insect pests that have developed resistance to existing insecticides.

SUMMARY

Compositions and methods for conferring pesticidal activity to bacteria,plants, plant cells, tissues and seeds are provided. Compositionsinclude nucleic acid molecules encoding sequences for pesticidal andinsecticidal polypeptides, vectors comprising those nucleic acidmolecules, and host cells comprising the vectors. Compositions alsoinclude the pesticidal polypeptide sequences and antibodies to thosepolypeptides. The nucleic acid sequences can be used in DNA constructsor expression cassettes for transformation and expression in organisms,including microorganisms and plants. The nucleotide or amino acidsequences may be synthetic sequences that have been designed forexpression in an organism including, but not limited to, a microorganismor a plant. Compositions also comprise transformed bacteria, plants,plant cells, tissues and seeds.

In particular, isolated or recombinant nucleic acid molecules areprovided encoding Pteridophyta Insecticidal Protein-50 (PtIP-50)polypeptides including amino acid substitutions, deletions, insertions,fragments thereof, and combinations thereof and PteridophytaInsecticidal Protein-65 (PtIP-65) polypeptides including amino acidsubstitutions, deletions, insertions, fragments thereof, andcombinations thereof. Additionally, amino acid sequences correspondingto the PtIP-50 polypeptides and PtIP-65 polypeptides are encompassed.Provided are isolated or recombinant nucleic acid molecules of SEQ IDNO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52,SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ IDNO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ ID NO: 70 capable ofencoding PtIP-50 polypeptides as well as amino acid substitutions,deletions, insertions, fragments thereof, and combinations thereof.Nucleic acid sequences that are complementary to a nucleic acid sequenceof the embodiments or that hybridize to a sequence of the embodimentsare also encompassed. Also provided are isolated or recombinant PtIP-50polypeptides of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO:74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ IDNO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88,SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO:93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ IDNO: 98 as well as amino acid substitutions, deletions, insertions,fragments thereof and combinations thereof. Provided are isolated orrecombinant nucleic acid molecules of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ IDNO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 or SEQ ID NO: 21 capable ofencoding PtIP-65 polypeptides as well as amino acid substitutions,deletions, insertions, fragments thereof, and combinations thereof.Nucleic acid sequences that are complementary to a nucleic acid sequenceof the embodiments or that hybridize to a sequence of the embodimentsare also encompassed. Also provided are isolated or recombinant PtIP-65polypeptides of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ IDNO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39,SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42 as well as amino acidsubstitutions, deletions, insertions, fragments thereof and combinationsthereof.

Methods are provided for producing the polypeptides and for using thosepolypeptides for controlling or killing a Lepidopteran, Coleopteran,nematode, fungi, and/or Dipteran pests. The transgenic plants of theembodiments express one or more of the pesticidal sequences disclosedherein. In various embodiments, the transgenic plant further comprisesone or more additional genes for insect resistance, for example, one ormore additional genes for controlling Coleopteran, Lepidopteran,Hemipteran or nematode pests. It will be understood by one of skill inthe art that the transgenic plant may comprise any gene imparting anagronomic trait of interest.

Methods for detecting the nucleic acids and polypeptides of theembodiments in a sample are also included. A kit for detecting thepresence of a PtIP-50 polypeptide and/or a PtIP-65 polypeptide ordetecting the presence of a polynucleotide encoding a PtIP-50polypeptide and/or a PtIP-65 polypeptide in a sample is provided. Thekit may be provided along with all reagents and control samplesnecessary for carrying out a method for detecting the intended agent, aswell as instructions for use.

The compositions and methods of the embodiments are useful for theproduction of organisms with enhanced pest resistance or tolerance.These organisms and compositions comprising the organisms are desirablefor agricultural purposes. The compositions of the embodiments are alsouseful for generating altered or improved proteins that have pesticidalactivity or for detecting the presence of PtIP-50 polypeptides ornucleic acids and PtIP-65 polypeptides or nucleic acids in products ororganisms.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 the phylogeny of ferns based on the classification for extantferns by A. R. Smith et al, TAXON, 55:705-731 (2006).

FIG. 2 shows the phylogeny tree of the PtIP-65 polypeptides: PtIP-65Aa(SEQ ID NO: 22), PtIP-65Ba (SEQ ID NO: 24), PtIP-65Bb (SEQ ID NO: 26),PtIP-65Ca (SEQ ID NO: 29), PtIP-65Fa (SEQ ID NO: 41), PtIP-65Fb (SEQ IDNO: 420, PtIP-65Ga (SEQ ID NO: 30), PtIP-65Gb (SEQ ID NO: 34), PtIP-65Gc(SEQ ID NO: 27), PtIP-65Gd (SEQ ID NO: 35), PtIP-65Ge (SEQ ID NO: 36),PtIP-65Ha (SEQ ID NO: 25), PtIP-65Hb (SEQ ID NO: 31), PtIP-65Hc (SEQ IDNO: 23), PtIP-65Hd (SEQ ID NO: 28), PtIP-65He (SEQ ID NO: 32), PtIP-65Hf(SEQ ID NO: 33), PtIP-65Hg (SEQ ID NO: 40), PtIP-65Hh (SEQ ID NO: 38),PtIP-65Hj (SEQ ID NO: 39), and PtIP-65Hk (SEQ ID NO: 37).

FIGS. 3a-3i show an alignment of the amino acid sequences of PtIP-65Aa(SEQ ID NO: 22), PtIP-65Ba (SEQ ID NO: 24), PtIP-65Bb (SEQ ID NO: 26),PtIP-65Ca (SEQ ID NO: 29), PtIP-65Fa (SEQ ID NO: 41), PtIP-65Fb (SEQ IDNO: 42), PtIP-65Ga (SEQ ID NO: 30), PtIP-65Gb (SEQ ID NO: 34), PtIP-65Gc(SEQ ID NO: 27), PtIP-65Gd (SEQ ID NO: 35), PtIP-65Ge (SEQ ID NO: 36),PtIP-65Ha (SEQ ID NO: 25), PtIP-65Hb (SEQ ID NO: 31), PtIP-65Hc (SEQ IDNO: 23), PtIP-65Hd (SEQ ID NO: 28), PtIP-65He (SEQ ID NO: 32), PtIP-65Hf(SEQ ID NO: 33), PtIP-65Hg (SEQ ID NO: 40), PtIP-65Hh (SEQ ID NO: 38),PtIP-65Hj (SEQ ID NO: 39), and PtIP-65Hk (SEQ ID NO: 37). The amino aciddiversity is indicated with shading.

FIGS. 4a-4b show an alignment of the amino acid sequences of PtIP-65Aa(SEQ ID NO: 22), PtIP-65Ba (SEQ ID NO: 24), PtIP-65Bb (SEQ ID NO: 26),and PtIP-65Ca (SEQ ID NO: 29), The amino acid diversity betweenPtIP-65Aa and PtIP-65Ba (SEQ ID NO: 24), PtIP-65Bb (SEQ ID NO: 26), andPtIP-65Ca is indicated with shading.

FIGS. 5a-5c show an alignment of the amino acid sequences of PtIP-65Ga(SEQ ID NO: 30), PtIP-65Gb (SEQ ID NO: 34), PtIP-65Gc (SEQ ID NO: 27),PtIP-65Gd (SEQ ID NO: 35), PtIP-65Ge (SEQ ID NO: 36), PtIP-65Ha (SEQ IDNO: 25). The amino acid diversity between PtIP-65Ga (SEQ ID NO: 30) andPtIP-65Gb (SEQ ID NO: 34), PtIP-65Gc (SEQ ID NO: 27), PtIP-65Gd (SEQ IDNO: 35), PtIP-65Ge (SEQ ID NO: 36), and PtIP-65Ha (SEQ ID NO: 25) isindicated with shading.

FIGS. 6a-6c show an alignment of the amino acid sequences of PtIP-65Hb(SEQ ID NO: 31), PtIP-65Hc (SEQ ID NO: 23), PtIP-65Hd (SEQ ID NO: 28),PtIP-65He (SEQ ID NO: 32), and PtIP-65Hf (SEQ ID NO: 33). The amino aciddiversity between PtIP-65Hb (SEQ ID NO: 31) and PtIP-65Hc (SEQ ID NO:23), PtIP-65Hd (SEQ ID NO: 28), PtIP-65He (SEQ ID NO: 32), and PtIP-65Hf(SEQ ID NO: 33) is indicated with shading.

FIG. 7 shows an alignment of the amino acid sequences of PtIP-65Fa (SEQID NO: 41) and PtIP-65Fb (SEQ ID NO: 42). The amino acid diversitybetween PtIP-65Fa (SEQ ID NO: 41) and PtIP-65Fb (SEQ ID NO: 42) isindicated with shading.

FIGS. 8a-8b show an alignment of the amino acid sequences of PtIP-65Hg(SEQ ID NO: 40) and PtIP-65Hh (SEQ ID NO: 38). The amino acid diversitybetween PtIP-65Hg (SEQ ID NO: 40) and PtIP-65Hh (SEQ ID NO: 38) isindicated with shading.

FIG. 9 shows the phylogeny tree of the PtIP-50 polypeptides: PtIP-50Aa(SEQ ID NO: 71), PtIP-50Ba (SEQ ID NO: 77), PtIP-50Bb (SEQ ID NO: 84),PtIP-50Bc (SEQ ID NO: 78), PtIP-50Bd (SEQ ID NO: 96), PtIP-50Fa (SEQ IDNO: 86), PtIP-50Fb (SEQ ID NO: 72), PtIP-50Fd (SEQ ID NO: 87), PtIP-50Fe(SEQ ID NO: 79), PtIP-50Ff (SEQ ID NO: 83), PtIP-50Fg (SEQ ID NO: 82),PtIP-50Fh (SEQ ID NO: 81), PtIP-50Fi (SEQ ID NO: 90), PtIP-50Fj (SEQ IDNO: 91), PtIP-50Fk (SEQ ID NO: 89), PtIP-50Fl (SEQ ID NO: 94), PtIP-50Fm(SEQ ID NO: 98), PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ ID NO: 73),PtIP-50Fp (SEQ ID NO: 97), PtIP-50Fq (SEQ ID NO: 95), PtIP-50Fr (SEQ IDNO: 75), PtIP-50Fs (SEQ ID NO: 74), PtIP-50Ft (SEQ ID NO: 85), PtIP-50Ga(SEQ ID NO: 88), PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQ ID NO: 76),and PtIP-50Gd (SEQ ID NO: 92).

FIGS. 10a-10o show an alignment of the amino acid sequences of PtIP-50Aa(SEQ ID NO: 71), PtIP-50Ba (SEQ ID NO: 77), PtIP-50Bb (SEQ ID NO: 84),PtIP-50Bc (SEQ ID NO: 78), PtIP-50Bd (SEQ ID NO: 96), PtIP-50Fa (SEQ IDNO: 86), PtIP-50Fb (SEQ ID NO: 72), PtIP-50Fd (SEQ ID NO: 87), PtIP-50Fe(SEQ ID NO: 79), PtIP-50Ff (SEQ ID NO: 83), PtIP-50Fg (SEQ ID NO: 82),PtIP-50Fh (SEQ ID NO: 81), PtIP-50Fi (SEQ ID NO: 90), PtIP-50Fj (SEQ IDNO: 91), PtIP-50Fk (SEQ ID NO: 89), PtIP-50Fl (SEQ ID NO: 94), PtIP-50Fm(SEQ ID NO: 98), PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ ID NO: 73),PtIP-50Fp (SEQ ID NO: 97), PtIP-50Fq (SEQ ID NO: 95), PtIP-50Fr (SEQ IDNO: 75), PtIP-50Fs (SEQ ID NO: 74), PtIP-50Ft (SEQ ID NO: 85), PtIP-50Ga(SEQ ID NO: 88), PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQ ID NO: 76),and PtIP-50Gd (SEQ ID NO: 92).

FIGS. 11a-11f show an alignment of the amino acid sequences of PtIP-50Aa(SEQ ID NO: 71), PtIP-50Ba (SEQ ID NO: 77), PtIP-50Bb (SEQ ID NO: 84),PtIP-50Bc (SEQ ID NO: 78), and PtIP-50Bd (SEQ ID NO: 96). The amino aciddiversity between PtIP-51Aa (SEQ ID NO: 71) and PtIP-50Ba (SEQ ID NO:77), PtIP-50Bb (SEQ ID NO: 84), PtIP-50Bc (SEQ ID NO: 78), and PtIP-50Bd(SEQ ID NO: 96) is indicated with shading.

FIGS. 12a-12j show an alignment of the amino acid sequences of PtIP-50Fb(SEQ ID NO: 72), PtIP-50Fe (SEQ ID NO: 79), PtIP-50Fk (SEQ ID NO: 89),PtIP-50Fl (SEQ ID NO: 94), PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ IDNO: 73), PtIP-50Fs (SEQ ID NO: 74), and PtIP-50Ft (SEQ ID NO: 85). Theamino acid diversity between PtIP-50Fb (SEQ ID NO: 72) and PtIP-50Fe(SEQ ID NO: 79), PtIP-50Fk (SEQ ID NO: 89), PtIP-50Fl (SEQ ID NO: 94),PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ ID NO: 73), PtIP-50Fs (SEQ IDNO: 74), and PtIP-50Ft (SEQ ID NO: 85) is indicated with shading.

FIGS. 13a-13e show an alignment of the amino acid sequences of PtIP-50Fd(SEQ ID NO: 87), PtIP-50Fg (SEQ ID NO: 82), PtIP-50Fh (SEQ ID NO: 81),PtIP-50Fm (SEQ ID NO: 98), and PtIP-50Fr (SEQ ID NO: 75). The amino aciddiversity between PtIP-50Fd (SEQ ID NO: 87) and PtIP-50Fg (SEQ ID NO:82), PtIP-50Fh (SEQ ID NO: 81), PtIP-50Fm (SEQ ID NO: 98), and PtIP-50Fr(SEQ ID NO: 75) is indicate with shading.

FIGS. 14a-14e show an alignment of the amino acid sequences of PtIP-50Ga(SEQ ID NO: 88), PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQ ID NO: 76),and PtIP-50Gd (SEQ ID NO: 92). The amino acid diversity betweenPtIP-50Ga (SEQ ID NO: 88) and PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQID NO: 76), and PtIP-50Gd (SEQ ID NO: 92) is indicated with shading.

FIGS. 15a-15e show an alignment of the amino acid sequences of PtIP-50Fa(SEQ ID NO: 86), PtIP-50Ff (SEQ ID NO: 83), PtIP-50Fi (SEQ ID NO: 90),PtIP-50Fj (SEQ ID NO: 91), and PtIP-50Fq (SEQ ID NO: 95). The amino aciddiversity between PtIP-50Fa (SEQ ID NO: 86) and PtIP-50Ff (SEQ ID NO:83), PtIP-50Fi (SEQ ID NO: 90), PtIP-50Fj (SEQ ID NO: 91), and PtIP-50Fq(SEQ ID NO: 95) is indicated with shading.

FIGS. 16a-16u show: an alignment of the amino acid sequences ofPtIP-50Aa (SEQ ID NO: 71), PtIP-50Ba (SEQ ID NO: 77), PtIP-50Bb (SEQ IDNO: 84), PtIP-50Bc (SEQ ID NO: 78), PtIP-50Bd (SEQ ID NO: 96), PtIP-50Fa(SEQ ID NO: 86), PtIP-50Fb (SEQ ID NO: 72), PtIP-50Fd (SEQ ID NO: 87),PtIP-50Fe (SEQ ID NO: 79), PtIP-50Ff (SEQ ID NO: 83), PtIP-50Fg (SEQ IDNO: 82), PtIP-50Fh (SEQ ID NO: 81), PtIP-50Fi (SEQ ID NO: 90), PtIP-50Fj(SEQ ID NO: 91), PtIP-50Fk (SEQ ID NO: 89), PtIP-50Fl (SEQ ID NO: 94),PtIP-50Fm (SEQ ID NO: 98), PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ IDNO: 73), PtIP-50Fp (SEQ ID NO: 97), PtIP-50Fq (SEQ ID NO: 95), PtIP-50Fr(SEQ ID NO: 75), PtIP-50Fs (SEQ ID NO: 74), PtIP-50Ft (SEQ ID NO: 85),PtIP-50Ga (SEQ ID NO: 88), PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQ IDNO: 76), and PtIP-50Gd (SEQ ID NO: 92); an alignment of the secondarystructure prediction for each of the PtIP-50 polypeptides, by thePSIPRED, top ranked secondary structure prediction method; and thelocations of the amino acid sequence motifs, as predicted by MEME motifanalysis, relative to PtIP-50Fb (SEQ ID NO: 72). A “H” indicates apredicted helical structure, an “E” indicates a PtIP-beta strandstructure, and a “C” indicates a predicted coil structure.

FIGS. 17a-17k show an alignment of the amino acid sequences of PtIP-65Aa(SEQ ID NO: 22), PtIP-65Ba (SEQ ID NO: 24), PtIP-65Bb (SEQ ID NO: 26),PtIP-65Ca (SEQ ID NO: 29), PtIP-65Fa (SEQ ID NO: 41), PtIP-65Fb (SEQ IDNO: 42), PtIP-65Ga (SEQ ID NO: 30), PtIP-65Gb (SEQ ID NO: 34), PtIP-65Gc(SEQ ID NO: 27), PtIP-65Gd (SEQ ID NO: 35), PtIP-65Ge (SEQ ID NO: 36),PtIP-65Ha (SEQ ID NO: 25), PtIP-65Hb (SEQ ID NO: 31), PtIP-65Hc (SEQ IDNO: 23), PtIP-65Hd (SEQ ID NO: 28), PtIP-65He (SEQ ID NO: 32), PtIP-65Hf(SEQ ID NO: 33), PtIP-65Hg (SEQ ID NO: 40), PtIP-65Hh (SEQ ID NO: 38),PtIP-65Hj (SEQ ID NO: 39), and PtIP-65Hk (SEQ ID NO: 37); an alignmentof the secondary structure prediction for each of the PtIP-65polypeptides, by the PSIPRED, top ranked secondary structure predictionmethod; and the locations of the amino acid sequence motifs, aspredicted by MEME motif analysis, relative to PtIP-65Gc (SEQ ID NO: 27).A “H” indicates a predicted helical structure, an “E” indicates aPtIP-beta strand structure, and a “C” indicates a predicted coilstructure.

FIG. 18 shows the effect of the PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa(SEQ ID NO: 22) polypeptides on isolated Anacarsia gemmatalis midgutactivity. I_(SCC) reflects the transport activity and midgut structuralintegrity to maintain normal midgut function. The decline in I_(SCC)following the addition of PtIP-50/65 polypeptides reflects the loss ofionic balance in the midgut.

FIG. 19 shows a gel image of the specific binding of PtIP-50Aa (SEQ IDNO: 71) and PtIP-65Aa (SEQ ID NO: 22) to BBMVs from C. includens.

FIGS. 20a and 20b show the evaluation of EC₅₀ values from densitometryof gel images for PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO:22) binding to C. includens BBMVs. FIG. 20a shows the averagedensitometry values for bound Alexa-PtIP-50Aa (SEQ ID NO: 71) in thepresence of different concentrations of unlabeled PtIP-50Aa (SEQ ID NO:71) and PtIP-65Aa (SEQ ID NO: 22) normalized to the amount bound in theabsence of unlabeled PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO:22). The solid line reflects the best fit of a square logistic equationto the data. FIG. 20b shows the average densitometry values for boundAlexa-PtIP-65Aa (SEQ ID NO: 22) in the presence of differentconcentrations of unlabeled PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQID NO: 22) normalized to the amount bound in the absence of unlabeledPtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22).

DETAILED DESCRIPTION

It is to be understood that this disclosure is not limited to theparticular methodology, protocols, cell lines, genera, and reagentsdescribed, as such may vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to limit the scope of the presentdisclosure.

As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof known to those skilled in the art, and so forth.All technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which thisdisclosure belongs unless clearly indicated otherwise.

The present disclosure is drawn to compositions and methods forcontrolling pests. The methods involve transforming organisms withnucleic acid sequences encoding PtIP-50 polypeptides and PtIP-65polypeptides. In particular, the nucleic acid sequences of theembodiments are useful for preparing plants and microorganisms thatpossess pesticidal activity. Thus, transformed bacteria, plants, plantcells, plant tissues and seeds are provided. The compositions arepesticidal nucleic acids and proteins of fern species. The nucleic acidsequences find use in the construction of expression vectors forsubsequent transformation into organisms of interest, as probes for theisolation of other homologous (or partially homologous) genes, and forthe generation of altered PtIP-50 polypeptides and PtIP-65 polypeptidesby methods known in the art, such as site directed mutagenesis, domainswapping or DNA shuffling. The PtIP-50 polypeptides and PtIP-65polypeptides find use in controlling or killing Lepidopteran,Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populationsand for producing compositions with pesticidal activity. Insect pests ofinterest include, but are not limited to, Lepidoptera species includingbut not limited to: Corn Earworm, (CEW) (Helicoverpa zea), European CornBorer (ECB) (Ostrinia nubialis), diamond-back moth, e.g., Helicoverpazea Boddie; soybean looper, e.g., Pseudoplusia includens Walker; andvelvet bean caterpillar e.g., Anticarsia gemmatalis Hübner andColeoptera species including but not limited to Western corn rootworm(Diabrotica virgifera)—WCRW, Southern corn rootworm (Diabroticaundecimpunctata howardi)—SCRW, and Northern corn rootworm (Diabroticabarberi)—NCRW.

By “pesticidal toxin” or “pesticidal protein” is used herein to refer toa toxin that has toxic activity against one or more pests, including,but not limited to, members of the Lepidoptera, Diptera, Hemiptera andColeoptera orders or the Nematoda phylum or a protein that has homologyto such a protein. Pesticidal proteins have been isolated from organismsincluding, for example, Bacillus sp., Pseudomonas sp., Photorhabdus sp.,Xenorhabdus sp., Clostridium bifermentans and Paenibacillus popilliae.Pesticidal proteins include but are not limited to: insecticidalproteins from Pseudomonas sp. such as PSEEN3174 (Monalysin; (2011) PLoSPathogens 7:1-13); from Pseudomonas protegens strain CHAO and Pf-5(previously fluorescens) (Pechy-Tarr, (2008) Environmental Microbiology10:2368-2386; GenBank Accession No. EU400157); from PseudomonasTaiwanensis (Liu, et al., (2010) J. Agric. Food Chem., 58:12343-12349)and from Pseudomonas pseudoalcligenes (Zhang, et al., (2009) Annals ofMicrobiology 59:45-50 and Li, et al., (2007) Plant Cell Tiss. OrganCult. 89:159-168); insecticidal proteins from Photorhabdus sp. andXenorhabdus sp. (Hinchliffe, et al., (2010) The Open Toxicology Journal,3:101-118 and Morgan, et al., (2001) Applied and Envir. Micro.67:2062-2069); U.S. Pat. Nos. 6,048,838, and 6,379,946; a PIP-1polypeptide of U.S. Ser. No. 13/792,861; an AfIP-1A and/or AfIP-1Bpolypeptide of U.S. Ser. No. 13/800,233; a PHI-4 polypeptide of U.S.Ser. No. 13/839,702; and δ-endotoxins including, but not limited to, theCry1, Cry2, Cry3, Cry4, Cry5, Cry6, Cry7, Cry8, Cry9, Cry10, Cry11,Cry12, Cry13, Cry14, Cry15, Cry16, Cry17, Cry18, Cry19, Cry20, Cry21,Cry22, Cry23, Cry24, Cry25, Cry26, Cry27, Cry 28, Cry 29, Cry 30, Cry31,Cry32, Cry33, Cry34, Cry35, Cry36, Cry37, Cry38, Cry39, Cry40, Cry41,Cry42, Cry43, Cry44, Cry45, Cry 46, Cry47, Cry49, Cry 51 and Cry55classes of 5-endotoxin genes and the B. thuringiensis cytolytic cyt1 andcyt2 genes. Members of these classes of B. thuringiensis insecticidalproteins include, but are not limited to Cry1Aa1 (Accession # AAA22353);Cry1Aa2 (Accession # Accession # AAA22552); Cry1Aa3 (Accession #BAA00257); Cry1Aa4 (Accession # CAA31886); Cry1Aa5 (Accession #BAA04468); Cry1Aa6 (Accession # AAA86265); Cry1Aa7 (Accession #AAD46139); Cry1Aa8 (Accession # I26149); Cry1Aa9 (Accession # BAA77213);Cry1Aa10 (Accession # AAD55382); Cry1Aa11 (Accession # CAA70856);Cry1Aa12 (Accession # AAP80146); Cry1Aa13 (Accession # AAM44305);Cry1Aa14 (Accession # AAP40639); Cry1Aa15 (Accession # AAY66993);Cry1Aa16 (Accession # HQ439776); Cry1Aa17 (Accession # HQ439788);Cry1Aa18 (Accession # HQ439790); Cry1Aa19 (Accession # HQ685121);Cry1Aa20 (Accession # JF340156); Cry1Aa21 (Accession # JN651496);Cry1Aa22 (Accession # KC158223); Cry1Ab1 (Accession # AAA22330); Cry1Ab2(Accession # AAA22613); Cry1Ab3 (Accession # AAA22561); Cry1Ab4(Accession # BAA00071); Cry1Ab5 (Accession # CAA28405); Cry1Ab6(Accession # AAA22420); Cry1Ab7 (Accession # CAA31620); Cry1Ab8(Accession # AAA22551); Cry1Ab9 (Accession # CAA38701); Cry1Ab10(Accession # A29125); Cry1Ab11 (Accession # I12419); Cry1Ab12 (Accession# AAC64003); Cry1Ab13 (Accession # AAN76494); Cry1Ab14 (Accession #AAG16877); Cry1Ab15 (Accession # AAO13302); Cry1Ab16 (Accession #AAK55546); Cry1Ab17 (Accession # AAT46415); Cry1Ab18 (Accession #AAQ88259); Cry1Ab19 (Accession # AAW31761); Cry1Ab20 (Accession #ABB72460); Cry1Ab21 (Accession # ABS18384); Cry1Ab22 (Accession #ABW87320); Cry1Ab23 (Accession # HQ439777); Cry1Ab24 (Accession #HQ439778); Cry1Ab25 (Accession # HQ685122); Cry1Ab26 (Accession #HQ847729); Cry1Ab27 (Accession # JN135249); Cry1Ab28 (Accession #JN135250); Cry1Ab29 (Accession # JN135251); Cry1Ab30 (Accession #JN135252); Cry1Ab31 (Accession # JN135253); Cry1Ab32 (Accession #JN135254); Cry1Ab33 (Accession # AAS93798); Cry1Ab34 (Accession #KC156668); Cry1Ab-like (Accession # AAK14336); Cry1Ab-like (Accession #AAK14337); Cry1Ab-like (Accession # AAK14338); Cry1Ab-like (Accession #ABG88858); Cry1Ac1 (Accession # AAA22331); Cry1Ac2 (Accession #AAA22338); Cry1Ac3 (Accession # CAA38098); Cry1Ac4 (Accession #AAA73077); Cry1Ac5 (Accession # AAA22339); Cry1Ac6 (Accession #AAA86266); Cry1Ac7 (Accession # AAB46989); Cry1Ac8 (Accession #AAC44841); Cry1Ac9 (Accession # AAB49768); Cry1Ac10 (Accession #CAA05505); Cry1Ac11 (Accession # CAA10270); Cry1Ac12 (Accession #I12418); Cry1Ac13 (Accession # AAD38701); Cry1Ac14 (Accession #AAQ06607); Cry1Ac15 (Accession # AAN07788); Cry1Ac16 (Accession #AAU87037); Cry1Ac17 (Accession # AAX18704); Cry1Ac18 (Accession #AAY88347); Cry1Ac19 (Accession # ABD37053); Cry1Ac20 (Accession #ABB89046); Cry1Ac21 (Accession # AAY66992); Cry1Ac22 (Accession #ABZ01836); Cry1Ac23 (Accession # CAQ30431); Cry1Ac24 (Accession #ABL01535); Cry1Ac25 (Accession # FJ513324); Cry1Ac26 (Accession #FJ617446); Cry1Ac27 (Accession # FJ617447); Cry1Ac28 (Accession #ACM90319); Cry1Ac29 (Accession # DQ438941); Cry1Ac30 (Accession #GQ227507); Cry1Ac31 (Accession # GU446674); Cry1Ac32 (Accession #HM061081); Cry1Ac33 (Accession # GQ866913); Cry1Ac34 (Accession #HQ230364); Cry1Ac35 (Accession # JF340157); Cry1Ac36 (Accession #JN387137); Cry1Ac37 (Accession # JQ317685); Cry1Ad1 (Accession #AAA22340); Cry1Ad2 (Accession # CAA01880); Cry1Ae1 (Accession #AAA22410); Cry1Af1 (Accession # AAB82749); Cry1Ag1 (Accession #AAD46137); Cry1Ah1 (Accession # AAQ14326); Cry1Ah2 (Accession #ABB76664); Cry1Ah3 (Accession # HQ439779); Cry1Ai1 (Accession #AAO39719); Cry1Ai2 (Accession # HQ439780); Cry1A-like (Accession #AAK14339); Cry1Ba1 (Accession # CAA29898); Cry1Ba2 (Accession #CAA65003); Cry1Ba3 (Accession # AAK63251); Cry1Ba4 (Accession #AAK51084); Cry1Ba5 (Accession # ABO20894); Cry1Ba6 (Accession #ABL60921); Cry1Ba7 (Accession # HQ439781); Cry1Bb1 (Accession #AAA22344); Cry1Bb2 (Accession # HQ439782); Cry1Bc1 (Accession #CAA86568); Cry1Bd1 (Accession # AAD10292); Cry1Bd2 (Accession #AAM93496); Cry1Be1 (Accession # AAC32850); Cry1Be2 (Accession #AAQ52387); Cry1Be3 (Accession # ACV96720); Cry1Be4 (Accession #HM070026); Cry1Bf1 (Accession # CAC50778); Cry1Bf2 (Accession #AAQ52380); Cry1Bg1 (Accession # AAO39720); Cry1Bh1 (Accession #HQ589331); Cry1Bi1 (Accession # KC156700); Cry1Ca1 (Accession #CAA30396); Cry1Ca2 (Accession # CAA31951); Cry1Ca3 (Accession #AAA22343); Cry1Ca4 (Accession # CAA01886); Cry1Ca5 (Accession #CAA65457); Cry1Ca6 [1] (Accession # AAF37224); Cry1Ca7 (Accession #AAG50438); Cry1Ca8 (Accession # AAM00264); Cry1Ca9 (Accession #AAL79362); Cry1Ca10 (Accession # AAN16462); Cry1Ca11 (Accession #AAX53094); Cry1Ca12 (Accession # HM070027); Cry1Ca13 (Accession #HQ412621); Cry1Ca14 (Accession # JN651493); Cry1Cb1 (Accession #M97880); Cry1Cb2 (Accession # AAG35409); Cry1Cb3 (Accession # ACD50894);Cry1Cb-like (Accession # AAX63901); Cry1Da1 (Accession # CAA38099);Cry1Da2 (Accession # I76415); Cry1Da3 (Accession # HQ439784); Cry1Db1(Accession # CAA80234); Cry1Db2 (Accession # AAK48937); Cry1Dc1(Accession # ABK35074); Cry1Ea1 (Accession # CAA37933); Cry1Ea2(Accession # CAA39609); Cry1Ea3 (Accession # AAA22345); Cry1Ea4(Accession # AAD04732); Cry1Ea5 (Accession # A15535); Cry1Ea6 (Accession# AAL50330); Cry1Ea7 (Accession # AAW72936); Cry1Ea8 (Accession #ABX11258); Cry1Ea9 (Accession # HQ439785); Cry1Ea10 (Accession #ADR00398); Cry1Ea11 (Accession # JQ652456); Cry1Eb1 (Accession #AAA22346); Cry1Fa1 (Accession # AAA22348); Cry1Fa2 (Accession #AAA22347); Cry1Fa3 (Accession # HM070028); Cry1Fa4 (Accession #HM439638); Cry1Fb1 (Accession # CAA80235); Cry1Fb2 (Accession #BAA25298); Cry1Fb3 (Accession # AAF21767); Cry1Fb4 (Accession #AAC10641); Cry1Fb5 (Accession # AAO13295); Cry1Fb6 (Accession #ACD50892); Cry1Fb7 (Accession # ACD50893); Cry1Ga1 (Accession #CAA80233); Cry1Ga2 (Accession # CAA70506); Cry1Gb1 (Accession #AAD10291); Cry1Gb2 (Accession # AAO13756); Cry1Gc1 (Accession #AAQ52381); Cry1Ha1 (Accession # CAA80236); Cry1Hb1 (Accession #AAA79694); Cry1Hb2 (Accession # HQ439786); Cry1H-like (Accession #AAF01213); Cry1Ia1 (Accession # CAA44633); Cry1Ia2 (Accession #AAA22354); Cry1Ia3 (Accession # AAC36999); Cry1Ia4 (Accession #AAB00958); Cry1Ia5 (Accession # CAA70124); Cry1Ia6 (Accession #AAC26910); Cry1Ia7 (Accession # AAM73516); Cry1Ia8 (Accession #AAK66742); Cry1Ia9 (Accession # AAQ08616); Cry1Ia10 (Accession #AAP86782); Cry1Ia11 (Accession # CAC85964); Cry1Ia12 (Accession #AAV53390); Cry1Ia13 (Accession # ABF83202); Cry1Ia14 (Accession #ACG63871); Cry1Ia15 (Accession # FJ617445); Cry1Ia16 (Accession #FJ617448); Cry1Ia17 (Accession # GU989199); Cry1Ia18 (Accession #ADK23801); Cry1Ia19 (Accession # HQ439787); Cry1Ia20 (Accession #JQ228426); Cry1Ia21 (Accession # JQ228424); Cry1Ia22 (Accession #JQ228427); Cry1Ia23 (Accession # JQ228428); Cry1Ia24 (Accession #JQ228429); Cry1Ia25 (Accession # JQ228430); Cry1Ia26 (Accession #JQ228431); Cry1Ia27 (Accession # JQ228432); Cry1Ia28 (Accession #JQ228433); Cry1Ia29 (Accession # JQ228434); Cry1Ia30 (Accession #JQ317686); Cry1Ia31 (Accession # JX944038); Cry1Ia32 (Accession #JX944039); Cry1Ia33 (Accession # JX944040); Cry1Ib1 (Accession #AAA82114); Cry1Ib2 (Accession # ABW88019); Cry1Ib3 (Accession #ACD75515); Cry1Ib4 (Accession # HM051227); Cry1Ib5 (Accession #HM070028); Cry1Ib6 (Accession # ADK38579); Cry1Ib7 (Accession #JN571740); Cry1Ib8 (Accession # JN675714); Cry1Ib9 (Accession #JN675715); Cry1Ib10 (Accession # JN675716); Cry1Ib11 (Accession #JQ228423); Cry1Ic1 (Accession # AAC62933); Cry1Ic2 (Accession #AAE71691); Cry1Id1 (Accession # AAD44366); Cry1Id2 (Accession #JQ228422); Cry1Ie1 (Accession # AAG43526); Cry1Ie2 (Accession #HM439636); Cry1Ie3 (Accession # KC156647); Cry1Ie4 (Accession #KC156681); Cry1If1 (Accession # AAQ52382); Cry1Ig1 (Accession #KC156701); Cry1I-like (Accession # AAC31094); Cry1I-like (Accession #ABG88859); Cry1Ja1 (Accession # AAA22341); Cry1Ja2 (Accession #HM070030); Cry1Ja3 (Accession # JQ228425); Cry1Jb1 (Accession #AAA98959); Cry1Jc1 (Accession # AAC31092); Cry1Jc2 (Accession #AAQ52372); Cry1Jd1 (Accession # CAC50779); Cry1Ka1 (Accession #AAB00376); Cry1Ka2 (Accession # HQ439783); Cry1La1 (Accession #AAS60191); Cry1La2 (Accession # HM070031); Cry1Ma1 (Accession #FJ884067); Cry1Ma2 (Accession # KC156659); Cry1Na1 (Accession #KC156648); Cry1Nb1 (Accession # KC156678); Cry1-like (Accession #AAC31091); Cry2Aa1 (Accession # AAA22335); Cry2Aa2 (Accession #AAA83516); Cry2Aa3 (Accession # D86064); Cry2Aa4 (Accession # AAC04867);Cry2Aa5 (Accession # CAA10671); Cry2Aa6 (Accession # CAA10672); Cry2Aa7(Accession # CAA10670); Cry2Aa8 (Accession # AAO13734); Cry2Aa9(Accession # AAO13750); Cry2Aa10 (Accession # AAQ04263); Cry2Aa11(Accession # AAQ52384); Cry2Aa12 (Accession # ABI83671); Cry2Aa13(Accession # ABL01536); Cry2Aa14 (Accession # ACF04939); Cry2Aa15(Accession # JN426947); Cry2Ab1 (Accession # AAA22342); Cry2Ab2(Accession # CAA39075); Cry2Ab3 (Accession # AAG36762); Cry2Ab4(Accession # AAO13296); Cry2Ab5 (Accession # AAQ04609); Cry2Ab6(Accession # AAP59457); Cry2Ab7 (Accession # AAZ66347); Cry2Ab8(Accession # ABC95996); Cry2Ab9 (Accession # ABC74968); Cry2Ab10(Accession # EF157306); Cry2Ab11 (Accession # CAM84575); Cry2Ab12(Accession # ABM21764); Cry2Ab13 (Accession # ACG76120); Cry2Ab14(Accession # ACG76121); Cry2Ab15 (Accession # HM037126); Cry2Ab16(Accession # GQ866914); Cry2Ab17 (Accession # HQ439789); Cry2Ab18(Accession # JN135255); Cry2Ab19 (Accession # JN135256); Cry2Ab20(Accession # JN135257); Cry2Ab21 (Accession # JN135258); Cry2Ab22(Accession # JN135259); Cry2Ab23 (Accession # JN135260); Cry2Ab24(Accession # JN135261); Cry2Ab25 (Accession # JN415485); Cry2Ab26(Accession # JN426946); Cry2Ab27 (Accession # JN415764); Cry2Ab28(Accession # JN651494); Cry2Ac1 (Accession # CAA40536); Cry2Ac2(Accession # AAG35410); Cry2Ac3 (Accession # AAQ52385); Cry2Ac4(Accession # ABC95997); Cry2Ac5 (Accession # ABC74969); Cry2Ac6(Accession # ABC74793); Cry2Ac7 (Accession # CAL18690); Cry2Ac8(Accession # CAM09325); Cry2Ac9 (Accession # CAM09326); Cry2Ac10(Accession # ABN15104); Cry2Ac11 (Accession # CAM83895); Cry2Ac12(Accession # CAM83896); Cry2Ad1 (Accession # AAF09583); Cry2Ad2(Accession # ABC86927); Cry2Ad3 (Accession # CAK29504); Cry2Ad4(Accession # CAM32331); Cry2Ad5 (Accession # CAO78739); Cry2Ae1(Accession # AAQ52362); Cry2Af1 (Accession # ABO30519); Cry2Af2(Accession # GQ866915); Cry2Ag1 (Accession # ACH91610); Cry2Ah1(Accession # EU939453); Cry2Ah2 (Accession # ACL80665); Cry2Ah3(Accession # GU073380); Cry2Ah4 (Accession # KC156702); Cry2Ai1(Accession # FJ788388); Cry2Aj (Accession #); Cry2Ak1 (Accession #KC156660); Cry2Ba1 (Accession # KC156658); Cry3Aa1 (Accession #AAA22336); Cry3Aa2 (Accession # AAA22541); Cry3Aa3 (Accession #CAA68482); Cry3Aa4 (Accession # AAA22542); Cry3Aa5 (Accession #AAA50255); Cry3Aa6 (Accession # AAC43266); Cry3Aa7 (Accession #CAB41411); Cry3Aa8 (Accession # AAS79487); Cry3Aa9 (Accession #AAW05659); Cry3Aa10 (Accession # AAU29411); Cry3Aa11 (Accession #AAW82872); Cry3Aa12 (Accession # ABY49136); Cry3Ba1 (Accession #CAA34983); Cry3Ba2 (Accession # CAA00645); Cry3Ba3 (Accession #JQ397327); Cry3Bb1 (Accession # AAA22334); Cry3Bb2 (Accession #AAA74198); Cry3Bb3 (Accession # I15475); Cry3Ca1 (Accession # CAA42469);Cry4Aa1 (Accession # CAA68485); Cry4Aa2 (Accession # BAA00179); Cry4Aa3(Accession # CAD30148); Cry4Aa4 (Accession # AFB18317); Cry4A-like(Accession # AAY96321); Cry4Ba1 (Accession # CAA30312); Cry4Ba2(Accession # CAA30114); Cry4Ba3 (Accession # AAA22337); Cry4Ba4(Accession # BAA00178); Cry4Ba5 (Accession # CAD30095); Cry4Ba-like(Accession # ABC47686); Cry4Ca1 (Accession # EU646202); Cry4Cb1(Accession # FJ403208); Cry4Cb2 (Accession # FJ597622); Cry4Cc1(Accession # FJ403207); Cry5Aa1 (Accession # AAA67694); Cry5Ab1(Accession # AAA67693); Cry5Ac1 (Accession # I34543); Cry5Ad1 (Accession# ABQ82087); Cry5Ba1 (Accession # AAA68598); Cry5Ba2 (Accession #ABW88931); Cry5Ba3 (Accession # AFJ04417); Cry5Ca1 (Accession #HM461869); Cry5Ca2 (Accession # ZP_04123426); Cry5Da1 (Accession #HM461870); Cry5Da2 (Accession # ZP_04123980); Cry5Ea1 (Accession #HM485580); Cry5Ea2 (Accession # ZP_04124038); Cry6Aa1 (Accession #AAA22357); Cry6Aa2 (Accession # AAM46849); Cry6Aa3 (Accession #ABH03377); Cry6Ba1 (Accession # AAA22358); Cry7Aa1 (Accession #AAA22351); Cry7Ab1 (Accession # AAA21120); Cry7Ab2 (Accession #AAA21121); Cry7Ab3 (Accession # ABX24522); Cry7Ab4 (Accession #EU380678); Cry7Ab5 (Accession # ABX79555); Cry7Ab6 (Accession #ACI44005); Cry7Ab7 (Accession # ADB89216); Cry7Ab8 (Accession #GU145299); Cry7Ab9 (Accession # ADD92572); Cry7Ba1 (Accession #ABB70817); Cry7Bb1 (Accession # KC156653); Cry7Ca1 (Accession #ABR67863); Cry7Cb1 (Accession # KC156698); Cry7Da1 (Accession #ACQ99547); Cry7Da2 (Accession # HM572236); Cry7Da3 (Accession #KC156679); Cry7Ea1 (Accession # HM035086); Cry7Ea2 (Accession #HM132124); Cry7Ea3 (Accession # EEM19403); Cry7Fa1 (Accession #HM035088); Cry7Fa2 (Accession # EEM19090); Cry7Fb1 (Accession #HM572235); Cry7Fb2 (Accession # KC156682); Cry7Ga1 (Accession #HM572237); Cry7Ga2 (Accession # KC156669); Cry7Gb1 (Accession #KC156650); Cry7Gc1 (Accession # KC156654); Cry7Gd1 (Accession #KC156697); Cry7Ha1 (Accession # KC156651); Cry71a1 (Accession #KC156665); Cry7Ja1 (Accession # KC156671); Cry7Ka1 (Accession #KC156680); Cry7Kb1 (Accession # BAM99306); Cry7La1 (Accession #BAM99307); Cry8Aa1 (Accession # AAA21117); Cry8Ab1 (Accession #EU044830); Cry8Ac1 (Accession # KC156662); Cry8Ad1 (Accession #KC156684); Cry8Ba1 (Accession # AAA21118); Cry8Bb1 (Accession #CAD57542); Cry8Bc1 (Accession # CAD57543); Cry8Ca1 (Accession #AAA21119); Cry8Ca2 (Accession # AAR98783); Cry8Ca3 (Accession #EU625349); Cry8Ca4 (Accession # ADB54826); Cry8Da1 (Accession #BAC07226); Cry8Da2 (Accession # BD133574); Cry8Da3 (Accession #BD133575); Cry8Db1 (Accession # BAF93483); Cry8Ea1 (Accession #AAQ73470); Cry8Ea2 (Accession # EU047597); Cry8Ea3 (Accession #KC855216); Cry8Fa1 (Accession # AAT48690); Cry8Fa2 (Accession #HQ174208); Cry8Fa3 (Accession # AFH78109); Cry8Ga1 (Accession #AAT46073); Cry8Ga2 (Accession # ABC42043); Cry8Ga3 (Accession #FJ198072); Cry8Ha1 (Accession # AAW81032); Cry81a1 (Accession #EU381044); Cry81a2 (Accession # GU073381); Cry81a3 (Accession #HM044664); Cry81a4 (Accession # KC156674); Cry81b1 (Accession #GU325772); Cry81b2 (Accession # KC156677); Cry8Ja1 (Accession #EU625348); Cry8Ka1 (Accession # FJ422558); Cry8Ka2 (Accession #ACN87262); Cry8Kb1 (Accession # HM123758); Cry8Kb2 (Accession #KC156675); Cry8La1 (Accession # GU325771); Cry8Ma1 (Accession #HM044665); Cry8Ma2 (Accession # EEM86551); Cry8Ma3 (Accession #HM210574); Cry8Na1 (Accession # HM640939); Cry8Pa1 (Accession #HQ388415); Cry8Qa1 (Accession # HQ441166); Cry8Qa2 (Accession #KC152468); Cry8Ra1 (Accession # AFP87548); Cry8Sa1 (Accession #JQ740599); Cry8Ta1 (Accession # KC156673); Cry8-like (Accession #FJ770571); Cry8-like (Accession # ABS53003); Cry9Aa1 (Accession #CAA41122); Cry9Aa2 (Accession # CAA41425); Cry9Aa3 (Accession #GQ249293); Cry9Aa4 (Accession # GQ249294); Cry9Aa5 (Accession #JX174110); Cry9Aa like (Accession # AAQ52376); Cry9Ba1 (Accession #CAA52927); Cry9Ba2 (Accession # GU299522); Cry9Bb1 (Accession #AAV28716); Cry9Ca1 (Accession # CAA85764); Cry9Ca2 (Accession #AAQ52375); Cry9Da1 (Accession # BAA19948); Cry9Da2 (Accession #AAB97923); Cry9Da3 (Accession # GQ249293); Cry9Da4 (Accession #GQ249297); Cry9Db1 (Accession # AAX78439); Cry9Dc1 (Accession #KC156683); Cry9Ea1 (Accession # BAA34908); Cry9Ea2 (Accession #AAO12908); Cry9Ea3 (Accession # ABM21765); Cry9Ea4 (Accession #ACE88267); Cry9Ea5 (Accession # ACF04743); Cry9Ea6 (Accession #ACG63872); Cry9Ea7 (Accession # FJ380927); Cry9Ea8 (Accession #GQ249292); Cry9Ea9 (Accession # JN651495); Cry9Eb1 (Accession #CAC50780); Cry9Eb2 (Accession # GQ249298); Cry9Eb3 (Accession #KC156646); Cry9Ec1 (Accession # AAC63366); Cry9Ed1 (Accession #AAX78440); Cry9Ee1 (Accession # GQ249296); Cry9Ee2 (Accession #KC156664); Cry9Fa1 (Accession # KC156692); Cry9Ga1 (Accession #KC156699); Cry9-like (Accession # AAC63366); Cry10Aa1 (Accession #AAA22614); Cry10Aa2 (Accession # E00614); Cry10Aa3 (Accession #CAD30098); Cry10Aa4 (Accession # AFB18318); Cry10A-like (Accession #DQ167578); Cry11Aa1 (Accession # AAA22352); Cry11Aa2 (Accession #AAA22611); Cry11Aa3 (Accession # CAD30081); Cry11Aa4 (Accession #AFB18319); Cry11Aa-like (Accession # DQ166531); Cry11Ba1 (Accession #CAA60504); Cry11Bb1 (Accession # AAC97162); Cry11Bb2 (Accession #HM068615); Cry12Aa1 (Accession # AAA22355); Cry13Aa1 (Accession #AAA22356); Cry14Aa1 (Accession # AAA21516); Cry14Ab1 (Accession #KC156652); Cry15Aa1 (Accession # AAA22333); Cry16Aa1 (Accession #CAA63860); Cry17Aa1 (Accession # CAA67841); Cry18Aa1 (Accession #CAA67506); Cry18Ba1 (Accession # AAF89667); Cry18Ca1 (Accession #AAF89668); Cry19Aa1 (Accession # CAA68875); Cry19Ba1 (Accession #BAA32397); Cry19Ca1 (Accession # AFM37572); Cry20Aa1 (Accession #AAB93476); Cry20Ba1 (Accession # ACS93601); Cry20Ba2 (Accession #KC156694); Cry20-like (Accession # GQ144333); Cry21Aa1 (Accession #I32932); Cry21Aa2 (Accession # I66477); Cry21Ba1 (Accession # BAC06484);Cry21Ca1 (Accession # JF521577); Cry21Ca2 (Accession # KC156687);Cry21Da1 (Accession # JF521578); Cry22Aa1 (Accession # I34547); Cry22Aa2(Accession # CAD43579); Cry22Aa3 (Accession # ACD93211); Cry22Ab1(Accession # AAK50456); Cry22Ab2 (Accession # CAD43577); Cry22Ba1(Accession # CAD43578); Cry22Bb1 (Accession # KC156672); Cry23Aa1(Accession # AAF76375); Cry24Aa1 (Accession # AAC61891); Cry24Ba1(Accession # BAD32657); Cry24Ca1 (Accession # CAJ43600); Cry25Aa1(Accession # AAC61892); Cry26Aa1 (Accession # AAD25075); Cry27Aa1(Accession # BAA82796); Cry28Aa1 (Accession # AAD24189); Cry28Aa2(Accession # AAG00235); Cry29Aa1 (Accession # CAC80985); Cry30Aa1(Accession # CAC80986); Cry30Ba1 (Accession # BAD00052); Cry30Ca1(Accession # BAD67157); Cry30Ca2 (Accession # ACU24781); Cry30Da1(Accession # EF095955); Cry30Db1 (Accession # BAE80088); Cry30Ea1(Accession # ACC95445); Cry30Ea2 (Accession # FJ499389); Cry30Fa1(Accession # ACI22625); Cry30Ga1 (Accession # ACG60020); Cry30Ga2(Accession # HQ638217); Cry31Aa1 (Accession # BAB11757); Cry31Aa2(Accession # AAL87458); Cry31Aa3 (Accession # BAE79808); Cry31Aa4(Accession # BAF32571); Cry31Aa5 (Accession # BAF32572); Cry31Aa6(Accession # BAI44026); Cry31Ab1 (Accession # BAE79809); Cry31Ab2(Accession # BAF32570); Cry31Ac1 (Accession # BAF34368); Cry31Ac2(Accession # AB731600); Cry31Ad1 (Accession # BAI44022); Cry32Aa1(Accession # AAG36711); Cry32Aa2 (Accession # GU063849); Cry32Ab1(Accession # GU063850); Cry32Ba1 (Accession # BAB78601); Cry32Ca1(Accession # BAB78602); Cry32Cb1 (Accession # KC156708); Cry32Da1(Accession # BAB78603); Cry32Ea1 (Accession # GU324274); Cry32Ea2(Accession # KC156686); Cry32Eb1 (Accession # KC156663); Cry32Fa1(Accession # K0156656); Cry32Ga1 (Accession # KC156657); Cry32Ha1(Accession # KC156661); Cry32Hb1 (Accession # KC156666); Cry32Ia1(Accession # KC156667); Cry32Ja1 (Accession # KC156685); Cry32Ka1(Accession # KC156688); Cry32La1 (Accession # K0156689); Cry32Ma1(Accession # KC156690); Cry32Mb1 (Accession # KC156704); Cry32Na1(Accession # KC156691); Cry32Oa1 (Accession # KC156703); Cry32Pa1(Accession # KC156705); Cry32Qa1 (Accession # KC156706); Cry32Ra1(Accession # K0156707); Cry32Sa1 (Accession # KC156709); Cry32Ta1(Accession # KC156710); Cry32Ua1 (Accession # KC156655); Cry33Aa1(Accession # AAL26871); Cry34Aa1 (Accession # AAG50341); Cry34Aa2(Accession # AAK64560); Cry34Aa3 (Accession # AAT29032); Cry34Aa4(Accession # AAT29030); Cry34Ab1 (Accession # AAG41671); Cry34Ac1(Accession # AAG50118); Cry34Ac2 (Accession # AAK64562); Cry34Ac3(Accession # AAT29029); Cry34Ba1 (Accession # AAK64565); Cry34Ba2(Accession # AAT29033); Cry34Ba3 (Accession # AAT29031); Cry35Aa1(Accession # AAG50342); Cry35Aa2 (Accession # AAK64561); Cry35Aa3(Accession # AAT29028); Cry35Aa4 (Accession # AAT29025); Cry35Ab1(Accession # AAG41672); Cry35Ab2 (Accession # AAK64563); Cry35Ab3(Accession # AY536891); Cry35Ac1 (Accession # AAG50117); Cry35Ba1(Accession # AAK64566); Cry35Ba2 (Accession # AAT29027); Cry35Ba3(Accession # AAT29026); Cry36Aa1 (Accession # AAK64558); Cry37Aa1(Accession # AAF76376); Cry38Aa1 (Accession # AAK64559); Cry39Aa1(Accession # BAB72016); Cry40Aa1 (Accession # BAB72018); Cry40Ba1(Accession # BAC77648); Cry40Ca1 (Accession # EU381045); Cry40Da1(Accession # ACF15199); Cry41Aa1 (Accession # BAD35157); Cry41Ab1(Accession # BAD35163); Cry41Ba1 (Accession # HM461871); Cry41Ba2(Accession # ZP_04099652); Cry42Aa1 (Accession # BAD35166); Cry43Aa1(Accession # BAD15301); Cry43Aa2 (Accession # BAD95474); Cry43Ba1(Accession # BAD15303); Cry43Ca1 (Accession # KC156676); Cry43Cb1(Accession # KC156695); Cry43Cc1 (Accession # KC156696); Cry43-like(Accession # BAD15305); Cry44Aa (Accession # BAD08532); Cry45Aa(Accession # BAD22577); Cry46Aa (Accession # BAC79010); Cry46Aa2(Accession # BAG68906); Cry46Ab (Accession # BAD35170); Cry47Aa(Accession # AAY24695); Cry48Aa (Accession # CAJ18351); Cry48Aa2(Accession # CAJ86545); Cry48Aa3 (Accession # CAJ86546); Cry48Ab(Accession # CAJ86548); Cry48Ab2 (Accession # CAJ86549); Cry49Aa(Accession # CAH56541); Cry49Aa2 (Accession # CAJ86541); Cry49Aa3(Accession # CAJ86543); Cry49Aa4 (Accession # CAJ86544); Cry49Ab1(Accession # CAJ86542); Cry50Aa1 (Accession # BAE86999); Cry50Ba1(Accession # GU446675); Cry50Ba2 (Accession # GU446676); Cry51Aa1(Accession # ABI14444); Cry51Aa2 (Accession # GU570697); Cry52Aa1(Accession # EF613489); Cry52Ba1 (Accession # FJ361760); Cry53Aa1(Accession # EF633476); Cry53Ab1 (Accession # FJ361759); Cry54Aa1(Accession # ACA52194); Cry54Aa2 (Accession # GQ140349); Cry54Ba1(Accession # GU446677); Cry55Aa1 (Accession # ABW88932); Cry54Ab1(Accession # JQ916908); Cry55Aa2 (Accession # AAE33526); Cry56Aa1(Accession # ACU57499); Cry56Aa2 (Accession # GQ483512); Cry56Aa3(Accession # JX025567); Cry57Aa1 (Accession # ANC87261); Cry58Aa1(Accession # ANC87260); Cry59Ba1 (Accession # JN790647); Cry59Aa1(Accession # ACR43758); Cry60Aa1 (Accession # ACU24782); Cry60Aa2(Accession # EAO57254); Cry60Aa3 (Accession # EEM99278); Cry60Ba1(Accession # GU810818); Cry60Ba2 (Accession # EAO57253); Cry60Ba3(Accession # EEM99279); Cry61Aa1 (Accession # HM035087); Cry61Aa2(Accession # HM132125); Cry61Aa3 (Accession # EEM19308); Cry62Aa1(Accession # HM054509); Cry63Aa1 (Accession # BAI44028); Cry64Aa1(Accession # BAJ05397); Cry65Aa1 (Accession # HM461868); Cry65Aa2(Accession # ZP_04123838); Cry66Aa1 (Accession # HM485581); Cry66Aa2(Accession # ZP_04099945); Cry67Aa1 (Accession # HM485582); Cry67Aa2(Accession # ZP_04148882); Cry68Aa1 (Accession # HQ113114); Cry69Aa1(Accession # HQ401006); Cry69Aa2 (Accession # JQ821388); Cry69Ab1(Accession # JN209957); Cry70Aa1 (Accession # JN646781); Cry70Ba1(Accession # ADO51070); Cry70Bb1 (Accession # EEL67276); Cry71Aa1(Accession # JX025568); Cry72Aa1 (Accession # JX025569); Cyt1Aa (GenBankAccession Number X03182); Cyt1Ab (GenBank Accession Number X98793);Cyt1B (GenBank Accession Number U37196); Cyt2A (GenBank Accession NumberZ14147); and Cyt2B (GenBank Accession Number U52043).

Examples of δ-endotoxins also include but are not limited to Cry1Aproteins of U.S. Pat. Nos. 5,880,275 and 7,858,849; a DIG-3 or DIG-11toxin (N-terminal deletion of α-helix 1 and/or α-helix 2 variants of cryproteins such as Cry1A, Cry3A) of U.S. Pat. Nos. 8,304,604, 8,304,605and 8,476,226; Cry1B of U.S. patent application Ser. No. 10/525,318;Cry1C of U.S. Pat. No. 6,033,874; Cry1F of U.S. Pat. Nos. 5,188,960 and6,218,188; Cry1A/F chimeras of U.S. Pat. Nos. 7,070,982; 6,962,705 and6,713,063); a Cry2 protein such as Cry2Ab protein of U.S. Pat. No.7,064,249); a Cry3A protein including but not limited to an engineeredhybrid insecticidal protein (eHIP) created by fusing unique combinationsof variable regions and conserved blocks of at least two different Cryproteins (US Patent Application Publication Number 2010/0017914); a Cry4protein; a Cry5 protein; a Cry6 protein; Cry8 proteins of U.S. Pat. Nos.7,329,736, 7,449,552, 7,803,943, 7,476,781, 7,105,332, 7,378,499 and7,462,760; a Cry9 protein such as such as members of the Cry9A, Cry9B,Cry9C, Cry9D, Cry9E and Cry9F families; a Cry15 protein of Naimov, etal., (2008) Applied and Environmental Microbiology, 74:7145-7151; aCry22, a Cry34Ab1 protein of U.S. Pat. Nos. 6,127,180, 6,624,145 and6,340,593; a CryET33 and cryET34 protein of U.S. Pat. Nos. 6,248,535,6,326,351, 6,399,330, 6,949,626, 7,385,107 and 7,504,229; a CryET33 andCryET34 homologs of US Patent Publication Number 2006/0191034,2012/0278954, and PCT Publication Number WO 2012/139004; a Cry35Ab1protein of U.S. Pat. Nos. 6,083,499, 6,548,291 and 6,340,593; a Cry46protein, a Cry 51 protein, a Cry binary toxin; a TIC901 or relatedtoxin; TIC807 of US Patent Application Publication Number 2008/0295207;ET29, ET37, TIC809, TIC810, TIC812, TIC127, TIC128 of PCT US2006/033867; AXMI-027, AXMI-036, and AXMI-038 of U.S. Pat. No.8,236,757; AXMI-031, AXMI-039, AXMI-040, AXMI-049 of U.S. Pat. No.7,923,602; AXMI-018, AXMI-020 and AXMI-021 of WO 2006/083891; AXMI-010of WO 2005/038032; AXMI-003 of WO 2005/021585; AXMI-008 of US PatentApplication Publication Number 2004/0250311; AXMI-006 of US PatentApplication Publication Number 2004/0216186; AXMI-007 of US PatentApplication Publication Number 2004/0210965; AXMI-009 of US PatentApplication Number 2004/0210964; AXMI-014 of US Patent ApplicationPublication Number 2004/0197917; AXMI-004 of US Patent ApplicationPublication Number 2004/0197916; AXMI-028 and AXMI-029 of WO2006/119457; AXMI-007, AXMI-008, AXMI-0080rf2, AXMI-009, AXMI-014 andAXMI-004 of WO 2004/074462; AXMI-150 of U.S. Pat. No. 8,084,416;AXMI-205 of US Patent Application Publication Number 2011/0023184;AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-019, AXMI-044, AXMI-037,AXMI-043, AXMI-033, AXMI-034, AXMI-022, AXMI-023, AXMI-041, AXMI-063 andAXMI-064 of US Patent Application Publication Number 2011/0263488;AXMI-R1 and related proteins of US Patent Application Publication Number2010/0197592; AXMI221Z, AXMI222z, AXMI223z, AXMI224z and AXMI225z of WO2011/103248; AXMI218, AXMI219, AXMI220, AXMI226, AXMI227, AXMI228,AXMI229, AXMI230 and AXMI231 of WO 2011/103247; AXMI-115, AXMI-113,AXMI-005, AXMI-163 and AXMI-184 of U.S. Pat. No. 8,334,431; AXMI-001,AXMI-002, AXMI-030, AXMI-035 and AXMI-045 of US Patent ApplicationPublication Number 2010/0298211; AXMI-066 and AXMI-076 of US PatentApplication Publication Number 2009/0144852; AXMI128, AXMI130, AXMI131,AXMI133, AXMI140, AXMI141, AXMI142, AXMI143, AXMI144, AXMI146, AXMI148,AXMI149, AXMI152, AXMI153, AXMI154, AXMI155, AXMI156, AXMI157, AXMI158,AXMI162, AXMI165, AXMI166, AXMI167, AXMI168, AXMI169, AXMI170, AXMI171,AXMI172, AXMI173, AXMI174, AXMI175, AXMI176, AXMI177, AXMI178, AXMI179,AXMI180, AXMI181, AXMI182, AXMI185, AXMI186, AXMI187, AXMI188, AXMI189of U.S. Pat. No. 8,318,900; AXMI079, AXMI080, AXMI081, AXMI082, AXMI091,AXMI092, AXMI096, AXMI097, AXMI098, AXMI099, AXMI100, AXMI101, AXMI102,AXMI103, AXMI104, AXMI107, AXMI108, AXMI109, AXMI110, AXMI111, AXMI112,AXMI114, AXMI116, AXMI117, AXMI118, AXMI119, AXMI120, AXMI121, AXMI122,AXMI123, AXMI124, AXMI1257, AXMI1268, AXMI127, AXMI129, AXMI164,AXMI151, AXMI161, AXMI183, AXMI132, AXMI138, AXMI137 of US PatentApplication Publication Number 2010/0005543, cry proteins such as Cry1Aand Cry3A having modified proteolytic sites of U.S. Pat. No. 8,319,019;a Cry1Ac, Cry2Aa and Cry1Ca toxin protein from Bacillus thuringiensisstrain VBTS 2528 of US Patent Application Publication Number2011/0064710. Other Cry proteins are well known to one skilled in theart (see, Crickmore, et al., “Bacillus thuringiensis toxin nomenclature”(2011), at lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/ which can beaccessed on the world-wide web using the “www” prefix). The insecticidalactivity of Cry proteins is well known to one skilled in the art (forreview, see, van Frannkenhuyzen, (2009) J. Invert. Path. 101:1-16). Theuse of Cry proteins as transgenic plant traits is well known to oneskilled in the art and Cry-transgenic plants including but not limitedto plants expressing Cry1Ac, Cry1Ac+Cry2Ab, Cry1Ab, Cry1A.105, Cry1F,Cry1Fa2, Cry1F+Cry1Ac, Cry2Ab, Cry3A, mCry3A, Cry3Bb1, Cry34Ab1,Cry35Ab1, Vip3A, mCry3A, Cry9c and CBI-Bt have received regulatoryapproval (see, Sanahuja, (2011) Plant Biotech Journal 9:283-300 and theCERA. (2010) GM Crop Database Center for Environmental Risk Assessment(CERA), ILSI Research Foundation, Washington D.C. atcera-gmc.org/index.php?action=gm_crop_database which can be accessed onthe world-wide web using the “www” prefix). More than one pesticidalproteins well known to one skilled in the art can also be expressed inplants such as Vip3Ab & Cry1Fa (US2012/0317682); Cry1BE & Cry1F(US2012/0311746); Cry1CA & Cry1AB (US2012/0311745); Cry1F & CryCa(US2012/0317681); Cry1DA & Cry1BE (US2012/0331590); Cry1DA & Cry1Fa(US2012/0331589); Cry1AB & Cry1BE (US2012/0324606); Cry1Fa & Cry2Aa andCry1I & Cry1E (US2012/0324605); Cry34Ab/35Ab and Cry6Aa (US20130167269);Cry34Ab/VCry35Ab & Cry3Aa (US20130167268); and Cry3A and Cry1Ab orVip3Aa (US20130116170). Pesticidal proteins also include insecticidallipases including lipid acyl hydrolases of U.S. Pat. No. 7,491,869, andcholesterol oxidases such as from Streptomyces (Purcell et al. (1993)Biochem Biophys Res Commun 15:1406-1413). Pesticidal proteins alsoinclude VIP (vegetative insecticidal proteins) toxins of U.S. Pat. Nos.5,877,012, 6,107,279 6,137,033, 7,244,820, 7,615,686, and 8,237,020 andthe like. Other VIP proteins are well known to one skilled in the art(see, lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/vip.html which can beaccessed on the world-wide web using the “www” prefix). Pesticidalproteins also include toxin complex (TC) proteins, obtainable fromorganisms such as Xenorhabdus, Photorhabdus and Paenibacillus (see, U.S.Pat. Nos. 7,491,698 and 8,084,418). Some TC proteins have “stand alone”insecticidal activity and other TC proteins enhance the activity of thestand-alone toxins produced by the same given organism. The toxicity ofa “stand-alone” TC protein (from Photorhabdus, Xenorhabdus orPaenibacillus, for example) can be enhanced by one or more TC protein“potentiators” derived from a source organism of a different genus.There are three main types of TC proteins. As referred to herein, ClassA proteins (“Protein A”) are stand-alone toxins. Class B proteins(“Protein B”) and Class C proteins (“Protein C”) enhance the toxicity ofClass A proteins. Examples of Class A proteins are TcbA, TcdA, XptA1 andXptA2. Examples of Class B proteins are TcaC, TcdB, XptB1Xb and XptC1Wi.Examples of Class C proteins are TccC, XptC1Xb and XptB1Wi. Pesticidalproteins also include spider, snake and scorpion venom proteins.Examples of spider venom peptides include but are not limited tolycotoxin-1 peptides and mutants thereof (U.S. Pat. No. 8,334,366).

In some embodiments the PtIP-50 polypeptide or the PtIP-65 polypeptideinclude amino acid sequences deduced from the full-length nucleic acidsequences disclosed herein and amino acid sequences that are shorterthan the full-length sequences, either due to the use of an alternatedownstream start site or due to processing that produces a shorterprotein having pesticidal activity. Processing may occur in the organismthe protein is expressed in or in the pest after ingestion of theprotein.

Thus, provided herein are novel isolated or recombinant nucleic acidsequences that confer pesticidal activity. Also provided are the aminoacid sequences of PtIP-50 polypeptides and PtIP-65 polypeptides. Theprotein resulting from translation of these PtIP-50 polypeptide and thePtIP-65 polypeptide genes allows cells to control or kill pests thatingest it.

Nucleic Acid Molecules, and Variants and Fragments Thereof

One aspect pertains to isolated or recombinant nucleic acid moleculescomprising nucleic acid sequences encoding PtIP-50 polypeptides, PtIP-65polypeptides or biologically active portions thereof, as well as nucleicacid molecules sufficient for use as hybridization probes to identifynucleic acid molecules encoding proteins with regions of sequencehomology. As used herein, the term “nucleic acid molecule” refers to DNAmolecules (e.g., recombinant DNA, cDNA, genomic DNA, plastid DNA,mitochondrial DNA) and RNA molecules (e.g., mRNA) and analogs of the DNAor RNA generated using nucleotide analogs. The nucleic acid molecule canbe single-stranded or double-stranded, but preferably is double-strandedDNA.

An “isolated” nucleic acid molecule (or DNA) is used herein to refer toa nucleic acid sequence (or DNA) that is no longer in its naturalenvironment, for example in vitro. A “recombinant” nucleic acid molecule(or DNA) is used herein to refer to a nucleic acid sequence (or DNA)that is in a recombinant bacterial or plant host cell. In someembodiments, an “isolated” or “recombinant” nucleic acid is free ofsequences (preferably protein encoding sequences) that naturally flankthe nucleic acid (i.e., sequences located at the 5′ and 3′ ends of thenucleic acid) in the genomic DNA of the organism from which the nucleicacid is derived. For purposes of the disclosure, “isolated” or“recombinant” when used to refer to nucleic acid molecules excludesisolated chromosomes. For example, in various embodiments, therecombinant nucleic acid molecule encoding PtIP-50 polypeptides orPtIP-65 polypeptides can contain less than about 5 kb, 4 kb, 3 kb, 2 kb,1 kb, 0.5 kb or 0.1 kb of nucleic acid sequences that naturally flankthe nucleic acid molecule in genomic DNA of the cell from which thenucleic acid is derived.

In some embodiments an isolated nucleic acid molecule encoding PtIP-50polypeptides or PtIP-65 polypeptides has one or more change in thenucleic acid sequence compared to the native or genomic nucleic acidsequence. In some embodiments the change in the native or genomicnucleic acid sequence includes but is not limited to: changes in thenucleic acid sequence due to the degeneracy of the genetic code; changesin the nucleic acid sequence due to the amino acid substitution,insertion, deletion and/or addition compared to the native or genomicsequence; removal of one or more intron; deletion of one or moreupstream or downstream regulatory regions; and deletion of the 5′ and/or3′ untranslated region associated with the genomic nucleic acidsequence. In some embodiments the nucleic acid molecule encoding aPtIP-50 polypeptide or PtIP-65 polypeptide is a non-genomic sequence.

A variety of polynucleotides that encode PtIP-50 polypeptides, PtIP-65polypeptides or related proteins are contemplated. Such polynucleotidesare useful for production of PtIP-50 polypeptides and PtIP-65polypeptides in host cells when operably linked to suitable promoter,transcription termination and/or polyadenylation sequences. Suchpolynucleotides are also useful as probes for isolating homologous orsubstantially homologous polynucleotides that encode PtIP-50polypeptides, PtIP-65 polypeptides or related proteins.

Polynucleotides Encoding PtIP-50 Polypeptides

One source of polynucleotides that encode PtIP-50 polypeptides orrelated proteins is a fern or other primitive plant species whichcontains a PtIP-50 polynucleotide of SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69 or SEQ ID NO: 70 encoding a PtIP-50 polypeptide of SEQ IDNO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80,SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO:85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ IDNO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98. Thepolynucleotides of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ IDNO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55,SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO:60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ IDNO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, andSEQ ID NO: 70 can be used to express PtIP-50 polypeptides in bacterialhosts that include but are not limited to Agrobacterium, Bacillus,Escherichia, Salmonella, Pseudomonas and Rhizobium bacterial host cells.The polynucleotides are also useful as probes for isolating homologousor substantially homologous polynucleotides that encode PtIP-50polypeptides or related proteins. Such probes can be used to identifyhomologous or substantially homologous polynucleotides derived fromPteridophyta species.

Polynucleotides that encode PtIP-50 polypeptides can also be synthesizedde novo from a PtIP-50 polypeptide sequence. The sequence of thepolynucleotide gene can be deduced from a PtIP-50 polypeptide sequencethrough use of the genetic code. Computer programs such as“BackTranslate” (GCG™ Package, Acclerys, Inc. San Diego, Calif.) can beused to convert a peptide sequence to the corresponding nucleotidesequence encoding the peptide. Examples of PtIP-50 polypeptide sequencesthat can be used to obtain corresponding nucleotide encoding sequencesinclude, but are not limited to the PtIP-50 polypeptides of SEQ ID NO:71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ IDNO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85,SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO:90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ IDNO: 95, SEQ ID NO: 96, SEQ ID NO: 97 and SEQ ID NO: 98. Furthermore,synthetic PtIP-50 polynucleotide sequences of the disclosure can bedesigned so that they will be expressed in plants. U.S. Pat. No.5,500,365 describes a method for synthesizing plant genes to improve theexpression level of the protein encoded by the synthesized gene. Thismethod relates to the modification of the structural gene sequences ofthe exogenous transgene, to cause them to be more efficientlytranscribed, processed, translated and expressed by the plant. Featuresof genes that are expressed well in plants include elimination ofsequences that can cause undesired intron splicing or polyadenylation inthe coding region of a gene transcript while retaining substantially theamino acid sequence of the toxic portion of the insecticidal protein. Asimilar method for obtaining enhanced expression of transgenes inmonocotyledonous plants is disclosed in U.S. Pat. No. 5,689,052.

In some embodiments the nucleic acid molecule encoding a PtIP-50polypeptide is a polynucleotide having the sequence set forth in SEQ IDNO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52,SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ IDNO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 and SEQ ID NO: 70, and variants,fragments and complements thereof. “Complement” is used herein to referto a nucleic acid sequence that is sufficiently complementary to a givennucleic acid sequence such that it can hybridize to the given nucleicacid sequence to thereby form a stable duplex. “Polynucleotide sequencevariants” is used herein to refer to a nucleic acid sequence that exceptfor the degeneracy of the genetic code encodes the same polypeptide.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide or PtIP-65 polypeptide is a non-genomic nucleic acidsequence. As used herein a “non-genomic nucleic acid sequence” or“non-genomic nucleic acid molecule” or “non-genomic polynucleotide”refers to a nucleic acid molecule that has one or more change in thenucleic acid sequence compared to a native or genomic nucleic acidsequence. In some embodiments the change to a native or genomic nucleicacid molecule includes but is not limited to: changes in the nucleicacid sequence due to the degeneracy of the genetic code; codonoptimization of the nucleic acid sequence for expression in plants;changes in the nucleic acid sequence to introduce at least one aminoacid substitution, insertion, deletion and/or addition compared to thenative or genomic sequence; removal of one or more intron associatedwith the genomic nucleic acid sequence; insertion of one or moreheterologous introns; deletion of one or more upstream or downstreamregulatory regions associated with the genomic nucleic acid sequence;insertion of one or more heterologous upstream or downstream regulatoryregions; deletion of the 5′ and/or 3′ untranslated region associatedwith the genomic nucleic acid sequence; insertion of a heterologous 5′and/or 3′ untranslated region; and modification of a polyadenylationsite. In some embodiments the non-genomic nucleic acid molecule is acDNA. In some embodiments the non-genomic nucleic acid molecule is asynthetic nucleic acid sequence.

In some embodiments the nucleic acid molecule encoding a PtIP-50polypeptide is a the non-genomic polynucleotide having a nucleotidesequence having at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%,59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%,73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%identity, to the nucleic acid sequence of SEQ ID NO: 43, SEQ ID NO: 44,SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO:49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ IDNO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63,SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO:68, SEQ ID NO: 69 or SEQ ID NO: 70, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO:98, wherein the PtIP-50 polypeptide, in combination with a PtIP-65polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 71, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 84or SEQ ID NO: 96, wherein the PtIP-50 polypeptide, in combination with aPtIP-65 polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 79,SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 93 or SEQ ID NO: 94, whereinthe PtIP-50 polypeptide, in combination with a PtIP-65 polypeptide, hasinsecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 75, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 87or SEQ ID NO: 98, wherein the PtIP-50 polypeptide, in combination with aPtIP-65 polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 76, SEQ ID NO: 80, SEQ ID NO: 88 or SEQ ID NO:92, wherein the PtIP-50 polypeptide, in combination with a PtIP-65polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 83, SEQ ID NO: 86, SEQ ID NO: 90 or SEQ ID NO:91, wherein the PtIP-50 polypeptide, in combination with a PtIP-65polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of SEQ ID NO: 97, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the any one of theamino acid sequences of SEQ ID NO: 224-326, wherein the PtIP-50polypeptide, in combination with a PtIP-65 polypeptide, has insecticidalactivity.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising an amino acid sequence of SEQ ID NO: 71, SEQ IDNO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81,SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO:86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ IDNO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98, having 1, 2, 3, 4, 5, 6, 7,8, 9, 10 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70 or more amino acid substitutions comparedto the native amino acid at the corresponding position of SEQ ID NO: 71,SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ IDNO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90,SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Division Pteridophyta.The phylogeny of ferns as used herein is based on the classification forextant ferns by A. R. Smith et al, TAXON, 55:705-731 (2006). Theconsensus phylogeny based on the classification by A. R. Smith is shownin FIG. 1. Other phylogenic classifications of extant ferns are known toone skilled in the art. Additional information on the phylogeny of fernscan be found at mobot.org/MOBOT/research/APweb/ (which can be accessedusing the “www” prefix) and Schuettpelz E. and Pryer K. M., TAXON 56:1037-1050 (2007) based on three plastid genes. Additional fern and otherprimitive plant species can be found athomepages.caverock.net.nz/˜byfern/list.htm (which can be accessed usingthe http:// prefix).

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Class Psilotopsida. Insome embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Class Psilotopsida,Order Psilotales. In some embodiments the nucleic acid molecule encodingPtIP-50 polypeptide is derived from a fern species in the ClassPsilotopsida, Order Ophioglossales. In some embodiments the nucleic acidmolecule encoding the PtIP-50 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales, Family Psilotaceae. Insome embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Class Psilotopsida,Order Ophioglossales Family Ophioglossaceae. In some embodiments thenucleic acid molecule encoding the PtIP-50 polypeptide is derived from afern species in the Genus Ophioglossum L., Botrychium, Botrypus,Helminthostachys, Ophioderma, Cheiroglossa, Sceptridium or Mankyua. Insome embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Ophioglossum L. Genusis selected from but not limited to Ophioglossum californicum,Ophioglossum coriaceum, Ophioglossum costatum, Ophioglossumcrotalophoroides, Ophioglossum engelmannii, Ophioglossum falcatum,Ophioglossum gomezianum, Ophioglossum gramineum, Ophioglossum kawamurae,Ophioglossum lusitanicum, Ophioglossum namegatae, Ophioglossumnudicaule, Ophioglossum palmatum, Ophioglossum parvum, Ophioglossumpedunculosum, Ophioglossum pendulum, Ophioglossum petiolatum,Ophioglossum pusillum, Ophioglossum reticulatum, Ophioglossumrichardsiae, Ophioglossum thermale, and Ophioglossum vulgatum.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the ClassPolypodiopsida/Pteridopsida. In some embodiments the nucleic acidmolecule encoding the PtIP-50 polypeptide is derived from a fern speciesin the Order Osmundales (royal ferns); Family Osmundaceae. In someembodiments the nucleic acid molecule encoding the PtIP-50 polypeptideis derived from a fern species in the Order Hymenophyllales (filmy fernsand bristle ferns); Family Hymenophyllaceae. In some embodiments thenucleic acid molecule encoding the PtIP-50 polypeptide is derived from afern species in the Order Gleicheniales; Family Gleicheniaceae, FamilyDipteridaceael or Family Matoniaceae. In some embodiments the nucleicacid molecule encoding the PtIP-50 polypeptide is derived from a fernspecies in the Order Schizaeales; Family Lygodiaceae, Family Anemiaceaeor Family Schizaeaceae. In some embodiments the nucleic acid moleculeencoding the PtIP-50 polypeptide is derived from a fern species in theOrder Salviniales; Family Marsileaceae or Family Salviniaceae. In someembodiments the nucleic acid molecule encoding the PtIP-50 polypeptideis derived from a fern species in the Order Cyatheales; FamilyThyrsopteridaceae, Family Loxsomataceae, Family Culcitaceae, FamilyPlagiogyriaceae, Family Cibotiaceae, Family Cyatheaceae, FamilyDicksoniaceae or Family Metaxyaceae.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales;Family Lindsaeaceae, Family Saccolomataceae, Family Cystodiaceae, FamilyDennstaedtiaceae, Family Pteridaceae, Family Aspleniaceae, FamilyThelypteridaceae, Family Woodsiaceae, Family Onocleaceae, FamilyBlechnaceae, Family Dryopteridaceae, Family Lomariopsidaceae, FamilyTectariaceae, Family Oleandraceae, Family Davalliaceae or FamilyPolypodiaceae.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Pteridaceae, Genus Adiantaceae selected from but not limited toAdiantum aethiopicum, Adiantum aleuticum, Adiantum bonatianum, Adiantumcajennense, Adiantum capillus-junonis, Adiantum capillus-veneris,Adiantum caudatum, Adiantum chienfi, Adiantum chilense, Adiantumcuneatum, Adiantum cunninghamfi, Adiantum davidii, Adiantum diaphanum,Adiantum edentulum, Adiantum edgeworthfi, Adiantum excisum, Adiantumfengianum, Adiantum fimbriatum, Adiantum flabellulatum, Adiantumformosanum, Adiantum formosum, Adiantum fulvum, Adiantum gravesfi,Adiantum hispidulum, Adiantum induratum, Adiantum jordanfi, Adiantumjuxtapositum, Adiantum latifolium, Adiantum leveillei, Adiantumlianxianense, Adiantum malesianum, Adiantum mariesfi, Adiantummonochlamys, Adiantum myriosorum, Adiantum obliquum, Adiantumogasawarense, Adiantum pedatum, Adiantum pentadactylon, Adiantumperuvianum, Adiantum philippense, Adiantum princeps, Adiantum pubescens,Adiantum raddianum, Adiantum reniforme, Adiantum roborowskii, Adiantumserratodentatum, Adiantum sinicum, Adiantum soboliferum, Adiantumsubcordatum, Adiantum tenerum, Adiantum terminatum, Adiantumtetraphyllum, Adiantum venustum, Adiantum viridescens, and Adiantumviridimontanum.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Aspleniaceae, Genus Asplenium L. In some embodiments the nucleicacid molecule encoding the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL selected from but not limited to Asplenium abbreviatum, Aspleniumabrotanoides, Asplenium abscissum var. subaequilaterale, Aspleniumabscissum, Asplenium achilleifolium, Asplenium acuminatum, Aspleniumadiantifrons, Asplenium adiantoides, Asplenium adiantoides var.squamulosum, Asplenium adiantum-nigrum L., Asplenium adiantum-nigrumvar. adiantum-nigrum, Asplenium adiantum-nigrum var. yuanurn, Aspleniumadnatum, Asplenium aethiopicum, Asplenium affine, Asplenium affine var.affine, Asplenium affine var. gilpinae, Asplenium affine var. mettenii,Asplenium affine var. pecten, Asplenium africanum, Asplenium afzelii,Asplenium aitchisonii, Asplenium alatulum, Asplenium alaturn, Aspleniumalfredii, Asplenium altajense, Asplenium amabile, Aspleniumambohitantelense, Asplenium anceps var. proliferum, Asplenium andapense,Asplenium andersonii, Asplenium angustatum, Asplenium angustum,Asplenium anisophyllum, Asplenium annetii, Asplenium antiquum, Aspleniumantrophyoides, Asplenium apertum, Asplenium apogamum, Aspleniumaquaticum, Asplenium arboreum, Asplenium arcanum, Asplenium arcuaturn,Asplenium argentinum, Asplenium argutum, Asplenium aspidiiforme,Asplenium aspidioides, Asplenium asterolepis, Asplenium auriculariumvar. acutidens, Asplenium auricularium var. subintegerrimum, Aspleniumauriculatum, Asplenium auriculatum var. aequilaterale, Asplenium auritumfo. diversifolium, Asplenium auritum fo. diversifolium, Aspleniumauritum fo. nana, Asplenium auritum, Asplenium auritum var. auriculatum,Asplenium auritum var. auritum, Asplenium auritum var. bipinnatifidum,Asplenium auritum var. bipinnatisectum, Asplenium auritum var.davallioides, Asplenium auritum var. macilentum, Asplenium auritum var.rigidum, Asplenium auritum var. subsimplex, Asplenium austrochinense,Asplenium ayopayense, Asplenium badinii, Asplenium balense, Aspleniumballivianii, Asplenium bangii, Asplenium bangii, Asplenium barbaense,Asplenium barclayanum, Asplenium barkamense, Asplenium barteri,Asplenium basiscopicum, Asplenium bicrenatum, Asplenium bifrons,Asplenium bipartitum, Asplenium blastophorum, Asplenium blepharodes,Asplenium blepharophorum, Asplenium boiteaui, Asplenium bolivianurn,Asplenium boltonii, Asplenium borealichinense, Asplenium bradei,Asplenium bradeorum, Asplenium bradleyi, Asplenium brausei, Aspleniumbreedlovei, Asplenium buettneri, Asplenium buettneri var. hildebrandtii,Asplenium bulbiferum, Asplenium bullatum var. bullatum, Aspleniumbullatum var. shikokianum, Asplenium bullatum, Asplenium cancellatum,Asplenium capillipes, Asplenium cardiophyllum (Hance), Aspleniumcaripense, Asplenium carvalhoanurn, Asplenium castaneoviride, Aspleniumcastaneum, Asplenium caudatum, Asplenium celtidifolium (Kunze),Asplenium ceratolepis, Asplenium changputungense, Asplenium chaseanum,Asplenium cheilosorum, Asplenium chengkouense, Asplenium chihuahuense,Asplenium chimantae, Asplenium chimborazense, Asplenium chingianurn,Asplenium chlorophyllum, Asplenium chondrophyllum, Asplenium cicutarium,Asplenium cicutarium var. paleaceum, Asplenium cirrhatum, Aspleniumcladolepton, Asplenium claussenii, Asplenium coenobiale, Aspleniumcommutatum, Asplenium congestum, Asplenium conquisitum, Aspleniumconsimile, Asplenium contiguum, Asplenium contiguum var. hirtulum,Asplenium corderoi, Asplenium cordovense, Asplenium coriaceum, Aspleniumcoriifolium, Asplenium correardii, Asplenium costale, Asplenium costalevar. robustum, Asplenium cowanii, Asplenium crenulatoserrulatum,Asplenium crenulatum, Asplenium crinicaule, Asplenium crinulosum,Asplenium cristatum, Asplenium cryptolepis Fernald, Aspleniumcultrifolium L., Asplenium cuneatiforme, Asplenium cuneaturn, Aspleniumcurvatura, Asplenium cuspidaturn, Asplenium cuspidatum var cuspidatum,Asplenium cuspidatum var. foeniculaceum, Asplenium cuspidatum var.triculum, Asplenium cuspidatum var. tripinnaturn, Asplenium dalhousiae,Asplenium dareoides, Asplenium davallioides, Asplenium davisii,Asplenium debile, Asplenium debile, Asplenium decussatum, Aspleniumdelavayi, Asplenium delicatulum, Asplenium delicatulum var. cocosensis,Asplenium delitescens, Asplenium delitescens×laetum, Asplenium densum,Asplenium dentatum L., Asplenium dentatum L., Asplenium depauperaturn,Asplenium deqenense, Asplenium dianae, Asplenium difforme, Aspleniumdilataturn, Asplenium dimidiaturn, Asplenium dimidiatum var. boliviense,Asplenium diplazisorum, Asplenium dissectum, Asplenium distans,Asplenium divaricatum, Asplenium divergens, Asplenium divisissimum,Asplenium doederleinii, Asplenium donnell-smithii, Asplenium dregeanum,Asplenium dulongjiangense, Asplenium duplicatoserratum, Aspleniumeatonii, Asplenium ebeneum, Asplenium ebenoides, Asplenium ecuadorense,Asplenium eggersii, Asplenium emarginatum, Asplenium enaturn, Aspleniumensiforme fo. bicuspe, Asplenium ensiforme fo. ensiforme, Aspleniumensiforme fo. stenophyllum, Asplenium ensiforme, Asplenium erectum var.erectum, Asplenium erectum var. gracile, Asplenium erectum var.usambarense, Asplenium erectum var. zeyheri, &, Asplenium erosum L.,Asplenium escaleroense, Asplenium esculentum, Asplenium eutecnum,Asplenium excelsum, Asplenium excisum, Asplenium exiguum, Aspleniumextensum, Asplenium falcaturn, Asplenium falcinellum, Asplenium faurei,Asplenium feel, Asplenium fengyangshanense, Asplenium ferulaceum,Asplenium fibrillosum, Asplenium filix-femina, Asplenium finckii,Asplenium finlaysonianurn, Asplenium flabellulatum, Aspleniumflabellulatum var flabellulatum, Asplenium flabellulatum var. partitum,Asplenium flaccidum, Asplenium flavescens, Asplenium flavidum, Aspleniumflexuosum, Asplenium fluminense, Asplenium foeniculaceum, Aspleniumformosanum, Asplenium formosum var. carolinurn, Asplenium formosum var.incultum, Asplenium formosum, Asplenium foumieri, Asplenium fragile,Asplenium fragile var. lomense, Asplenium fragrans, Asplenium fragransvar. foeniculaceum, Asplenium franconis var. gracile, Aspleniumfraxinifolium, Asplenium friesiorum, Asplenium friesiorum var.nesophilum, Asplenium fugax, Asplenium fujianense, Asplenium furcatum,Asplenium furfuraceum, Asplenium fuscipes, Asplenium fuscopubescens,Asplenium galeottii, Asplenium gautieri, Asplenium gemmiferum, Aspleniumgentryi, Asplenium geppii, Asplenium ghiesbreghtii, Asplenium gilliesii,Asplenium gilpinae, Asplenium glanduliserraturn, Asplenium glenniei,Asplenium goldmannii, Asplenium gomezianurn, Asplenium grande, Aspleniumgrandifolium, Asplenium grandifrons, Asplenium gregoriae, Aspleniumgriffithianum, Asplenium gulingense, Asplenium hainanense, Aspleniumhallbergii, Asplenium hallei, Asplenium hallii, Asplenium hangzhouense,Asplenium haplophyllum, Asplenium harpeodes, Asplenium harpeodes var.glaucovirens, Asplenium harpeodes var. incisura, Asplenium harrisiiJenman, Asplenium harrisonii, Asplenium hastatum, Asplenium hebeiense,Asplenium hemionitideum, Asplenium hemitomurn, Asplenium henryi,Asplenium herpetopteris, Asplenium herpetopteris var herpetopteris,Asplenium herpetopteris var. acutipinnata, Asplenium herpetopteris var.masoulae, Asplenium herpetopteris var. villosum, Asplenium hesperium,Asplenium heterochroum, Asplenium hians, Asplenium hians var.pallescens, Asplenium hoffmannii, Asplenium holophlebium, Aspleniumhondoense, Asplenium horridum, Asplenium hostmannii, Aspleniumhumistratum, Asplenium hypomelas, Asplenium inaequilaterale, Aspleniumincisura, Asplenium incurvaturn, Asplenium indicum, Asplenium indicumvar. indicum, Asplenium indicum var. yoshingagae, Asplenium induratum,Asplenium indusiatum, Asplenium inexpectatum, Asplenium insigne,Asplenium insiticium, Asplenium insolitum, Asplenium integerrimurn,Asplenium interjectum, Asplenium jamesonii, Asplenium jaundeense,Asplenium juglandifolium, Asplenium kangdingense, Asplenium kansuense,Asplenium kassneri, Asplenium kaulfussii, Asplenium kellermanii,Asplenium kentuckiense, Asplenium khullarii, Asplenium kiangsuense,Asplenium kunzeanurn, Asplenium lacerum, Asplenium laciniatum, Aspleniumlaciniatum var. acutipinna, Asplenium laciniatum var. laciniatum,Asplenium laetum fo. minor, Asplenium laetum, Asplenium laetum var.incisoserratum, Asplenium lamprocaulon, Asplenium laserpitiifolium var.morrisonense, Asplenium lastii, Asplenium latedens, Aspleniumlatifolium, Asplenium laui, Asplenium laurentii, Asplenium leandrianurn,Asplenium lechleri, Asplenium leiboense, Asplenium lepidorachis,Asplenium leptochlamys, Asplenium leptophyllum, Asplenium levyi,Asplenium lindbergii, Asplenium lindeni, Asplenium lineatum, Aspleniumlividum, Asplenium lobatum, Asplenium lobulaturn, Asplenium lokohoense,Asplenium longicauda, Asplenium longicaudaturn, Asplenium longifolium,Asplenium longisorum, Asplenium longjinense, Asplenium lorentzii,Asplenium loriceum, Asplenium loxogrammoides, Asplenium lugubre,Asplenium lunulatum, Asplenium lunulatum var. pteropus, Aspleniumlushanense, Asplenium lydgatei, Asplenium macilentum, Asplenium macraei,Asplenium macrodictyon, Asplenium macrophlebium, Asplenium macrophyllum,Asplenium macropterum, Asplenium macrosorum, Asplenium macrotis,Asplenium macrurum, Asplenium mainlingense, Asplenium mangindranense,Asplenium mannii, Asplenium marginatum L., Asplenium marojejyense,Asplenium martianum, Asplenium matsumurae, Asplenium mauritiensisLorence, Asplenium maximum, Asplenium, ii, Asplenium megalura, Aspleniummegaphyllum, Asplenium meiotomum, Asplenium melanopus, Aspleniummembranifolium, Asplenium meniscioides, Asplenium mesosorum, Aspleniummexicanum, Asplenium micropaleatum, Asplenium microtum, Aspleniummildbraedii, Asplenium mildei, Asplenium minimum, Asplenium minutum,Asplenium miradorense, Asplenium miyunense, Asplenium moccenianum,Asplenium mocquetysii, Asplenium modestum, Asplenium monanthemum var.menziesii, Asplenium monanthes L., Asplenium monanthes var monanthes,Asplenium monanthes var. castaneum, Asplenium monanthes var. wagneri,Asplenium monanthes var. yungense, Asplenium monodon, Aspleniummontanum, Asplenium mosetenense, Asplenium moupinense, Aspleniummucronatum, Asplenium munchii, Asplenium muticum, Asplenium myapteron,Asplenium myriophyllu, Asplenium nakanoanum, Asplenium nanchuanense,Asplenium nemorale, Asplenium neolaserpitiifolium, Aspleniumneomutijugum, Asplenium neovarians, Asplenium nesii, Aspleniumnesioticum, Asplenium nidus L., Asplenium nigricans, Aspleniumniponicum, Asplenium normale, Asplenium normale var. angustum, Aspleniumobesum, Asplenium oblongatum, Asplenium oblongifolium, Aspleniumobovatum, Asplenium obscurum, Asplenium obscurum var. angustum,Asplenium obtusatum var. obtusatum, Asplenium obtusatum var. sphenoides,Asplenium obtusifolium L., Asplenium obtusissimum, Asplenium obversum,Asplenium ochraceum, Asplenium oellgaardii, Asplenium ofeliae, Aspleniumoldhami, Asplenium oligosorum, Asplenium olivaceum, Asplenium onopterisL., Asplenium onustum, Asplenium ortegae, Asplenium otites, Aspleniumpalaciosii, Asplenium palmeri, Asplenium partitum, Asplenium parvisorum,Asplenium parviusculum, Asplenium parvulum, Asplenium patens, Aspleniumpaucifolium, Asplenium paucijugum, Asplenium paucivenosum, Aspleniumpearcei, Asplenium pekinense, Asplenium pellucidum, Asplenium pendulum,Asplenium petiolulatum, Asplenium phyllitidis, Aspleniumpimpinellifolium, Asplenium pinnatifidum, Asplenium pinnatum, Aspleniumplatyneuron, Asplenium platyneuron var. bacculum-rubrum, Aspleniumplatyneuron var. incisum, Asplenium platyphyllum, Asplenium plumbeum,Asplenium poloense, Asplenium polymeris, Asplenium polymorphum,Asplenium polyodon, Asplenium polyodon var. knudsenii, Aspleniumpolyodon var. nitidulum, Asplenium polyodon var. sectum, Aspleniumpolyodon var. subcaudatum, Asplenium polyphyllum, Asplenium poolii,Asplenium poolii fo. simplex, Asplenium poolii var. linearipinnatum,Asplenium potosinum, Asplenium potosinum var. incisum, Aspleniumpraegracile, Asplenium praemorsum, Asplenium preussii, Aspleniumpringleanum, Asplenium pringlei, Asplenium prionitis, Aspleniumprocerum, Asplenium progrediens, Asplenium projectum, Aspleniumprolongatum, Asplenium propinquum, Asplenium protensum, Aspleniumpseudoangustum, Asplenium pseudoerectum, Asplenium pseudofontanum,Asplenium pseudolaserpitiifolium, Asplenium pseudonormale, Aspleniumpseudo pellucidum, Asplenium pseudopraemorsum, Asplenium pseudovarians,Asplenium pseudowilfordii, Asplenium pseudowrightii, Aspleniumpsilacrum, Asplenium pteropus, Asplenium pubirhizoma, Aspleniumpulchellum, Asplenium pulchellum var. subhorizontale, Aspleniumpulcherrimum, Asplenium pulicosum, Asplenium pulicosum var. maius,Asplenium pululahuae, Asplenium pumilum, Asplenium pumilum var.hymenophylloides, Asplenium pumilum var. laciniatum, Aspleniumpurdieanum, Asplenium purpurascens, Asplenium pyramidatum, Aspleniumqiujiangense, Asplenium quercicola, Asplenium quitense, Aspleniumraddianum, Asplenium radiatum, Asplenium radicans L., Aspleniumradicans, Asplenium radicans var. costaricense, Asplenium radicans var.partitum, Asplenium radicans var. radicans, Asplenium radicans var.uniseriale, Asplenium recumbens, Asplenium reflexum, Asplenium regularevar. latior, Asplenium repandulum, Asplenium repens, Asplenium repente,Asplenium resiliens, Asplenium retusulum, Asplenium rhipidoneuron,Asplenium rhizophorum L., Asplenium rhizophyllum, Asplenium rhizophyllumL., Asplenium rhizophyllum var. proliferum, Asplenium rhomboideum,Asplenium rigidum, Asplenium riparium, Asplenium rivale, Aspleniumrockii, Asplenium roemerianum, Asplenium roemerianum var. mindensis,Asplenium rosenstockianum, Asplenium rubinum, Asplenium ruizianum,Asplenium rusbyanum, Asplenium ruta-muraria L., Asplenium ruta-murariavar. cryptolepis, Asplenium rutaceum, Asplenium rutaceum var.disculiferum, Asplenium rutaefolium, Asplenium rutifolium, Aspleniumsalicifolium L., Asplenium salicifolium var. aequilaterale, Aspleniumsalicifolium var. salicifolium, Asplenium sampsoni, Asplenium sanchezii,Asplenium sanderi, Asplenium sandersonii, Asplenium sanguinolentum,Asplenium sarelii, Asplenium sarelii var. magnum, Asplenium sarelii var.sarelii, Asplenium saxicola, Asplenium scalifolium, Aspleniumscandicinum, Asplenium schizophyllum, Asplenium schkuhrii, Aspleniumsciadophilum, Asplenium scolopendrium L., Asplenium scortechinii,Asplenium seileri, Asplenium semipinnatum, Asplenium septentrionale,Asplenium serra, Asplenium serra var. imrayanum, Aspleniumserratissimum, Asplenium serratum L., Asplenium serratum var. caudatum,Asplenium serricula, Asplenium sessilifolium, Asplenium sessilifoliumvar. guatemalense, Asplenium sessilifolium var. minus, Aspleniumsessilifolium var. occidentale, Asplenium sessilipinnum, Aspleniumsetosum, Asplenium shepherdii, Asplenium shepherdii var. bipinnatum,Asplenium shepherdii var. flagelliferum, Asplenium shikokianum,Asplenium simii, Asplenium simonsianum, Asplenium sintenisii, Aspleniumskinneri, Asplenium skinneri, Asplenium sodiroi, Aspleniumsoleirolioides, Asplenium solidum var. stenophyllum, Asplenium solmsii,Asplenium sp.-N.-Halle-2234, Asplenium spathulinum, Aspleniumspectabile, Asplenium speluncae, Asplenium sphaerosporum, Aspleniumsphenotomurn, Asplenium spinescens, Asplenium splendens, Aspleniumsprucei, Asplenium squamosum L., Asplenium standleyi, Aspleniumstellatum, Asplenium stenocarpum, Asplenium stoloniferum, Aspleniumstolonipes, Asplenium striatum L., Asplenium stuebelianum, Aspleniumstuhlmannii, Asplenium suave, Asplenium subalatum, Aspleniumsubcrenaturn, Asplenium subdigitatum, Asplenium subdimidiatum, Aspleniumsubintegrum, Asplenium sublaserpitfifolium, Asplenium sublongum,Asplenium subnudum, Asplenium suborbiculare, Asplenium subtenuifolium,Asplenium subtile, Asplenium subtoramanurn, Asplenium subtrapezoideum,Asplenium subvarians, Asplenium sulcatum, Asplenium sylvaticum,Asplenium szechuanense, Asplenium taiwanense, Asplenium tenerrimum,Asplenium tenerum, Asplenium tenuicaule, Asplenium tenuifolium,Asplenium tenuifolium var. minor, Asplenium tenuifolium var.tenuifolium, Asplenium tenuissimum, Asplenium ternatum, Aspleniumtheciferum, Asplenium theciferum var. concinnum, Asplenium thunbergii,Asplenium tianmushanense, Asplenium tianshanense, Asplenium tibeticum,Asplenium tocoraniense, Asplenium toramanurn, Asplenium trapezoideum,Asplenium tricholepis, Asplenium trichomanes L., Asplenium trichomanessubsp. inexpectans, Asplenium trichomanes subsp. quadrivalens, Aspleniumtrichomanes subsp. trichomanes, Asplenium trichomanes var. harovii,Asplenium trichomanes var. herbaceum, Asplenium trichomanes var. repens,Asplenium trichomanes var. viridissimum, Asplenium trichomanes-dentatumL., Asplenium trigonopterum, Asplenium trilobatum, Asplenium trilobum,Asplenium triphyllum, Asplenium triphyllum var. cornpactum, Aspleniumtriphyllum var. gracillimum, Asplenium triphyllum var. herbaceum,Asplenium tripteropus, Asplenium triquetrum, Asplenium truncorum,Asplenium tsaratananense, Asplenium tucumanense, Asplenium tuerckheimii,Asplenium tunquiniense, Asplenium ulbrichtii, Asplenium ultimurn,Asplenium unilaterale, Asplenium unilaterale var. decurrens, Aspleniumunilaterale var. udum, Asplenium unilaterale var. unilaterale, Aspleniumuniseriale, Asplenium uropteron, Asplenium vegans, Aspleniumvareschianum, Asplenium variabile var. paucijugum, Asplenium variabilevar. variabile, Asplenium varians subsp. fimbriatum, Asplenium varians,Asplenium vastum, Asplenium venturae, Asplenium venulosum, Aspleniumverapax, Asplenium vesiculosum, Asplenium vespertinurn, Aspleniumvillosum, Asplenium virens, Asplenium viride, Asplenium viridifrons,Asplenium virillae, Asplenium viviparioides, Asplenium viviparum,Asplenium viviparum var viviparum, Asplenium viviparum var. lineatu,Asplenium volubile, Asplenium vulcanicum, Asplenium wacketii, Aspleniumwagneri, Asplenium wallichianum, Asplenium wameckei, Aspleniumwilfordii, Asplenium williamsii, Asplenium wrightii, Aspleniumwrightioides, Asplenium wuliangshanense, Asplenium xianqianense,Asplenium xinjiangense, Asplenium xinyiense, Asplenium yelagagense,Asplenium yoshinagae, Asplenium yunnanense, Asplenium zamiifolium,Asplenium zanzibaricum, Asplenium biscayneanum, Asplenium curtissii,Asplenium ebenoides, Asplenium herb-wagneri, Asplenium heteroresiliens,Asplenium kenzoi, Asplenium plenum, Asplenium wangii, andAsplenium×clermontiae, Asplenium×gravesii.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Blechnaceae, Genus Blechnum L. In some embodiments the nucleicacid molecule encoding the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.selected from but not limited to Blechnum amabile, Blechnumappendiculatum, Blechnum articulatum, Blechnum australe, Blechnumaustrobrasilianum, Blechnum binervatum, Blechnum blechnoides, Blechnumbrasiliense, Blechnum capense, Blechnum cartilagineum, Blechnumcastaneum, Blechnum chambersii, Blechnum chilense, Blechnum colensoi,Blechnum contiguum, Blechnum cordatum, Blechnum coriaceum, Blechnumdiscolor, Blechnum doodioides, Blechnum durum, Blechnum eburneum,Blechnum ensiforme, Blechnum filiforme, Blechnum fluviatile, Blechnumfragile, Blechnum fraseri, Blechnum fullagari, Blechnum gibbum, Blechnumglandulosum, Blechnum gracile, Blechnum hancockii, Blechnum hastatum,Blechnum howeanum, Blechnum indicum, Blechnum kunthianum, Blechnumlaevigatum, Blechnum loxense, Blechnum magellanicum, Blechnummembranaceum, Blechnum microbasis, Blechnum microphyllum, Blechnummilnei, Blechnum minus, Blechnum mochaenum, Blechnum montanum, Blechnummoorei, Blechnum moritzianum, Blechnum nigrum, Blechnum niponicum,Blechnum norfolkianum, Blechnum novae-zelandiae, Blechnum nudum,Blechnum obtusatum, Blechnum occidentale, Blechnum oceanicum, Blechnumorientale, Blechnum patersonii, Blechnum penna-marina, Blechnumpolypodioides, Blechnum procerum, Blechnum punctulatum, Blechnumsampaioanum, Blechnum schiedeanum, Blechnum schomburgkii, Blechnumserrulatum, Blechnum simillimum, Blechnum spicant, Blechnumstipitellatum, Blechnum tabulare, Blechnum triangularifolium, Blechnumvieillardii, Blechnum vulcanicum, Blechnum wattsii, Blechnum whelanii,and Blechnum wurunuran.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Dryopteridaceae Genus Acrophorus, Genus Acrorumohra, GenusAnapausia, Genus Arachniodes, Genus Bolbitis, Genus Ctenitis, GenusCyclodium, Genus Cyrtogonellum, Genus Cyrtomidictyum, Genus Cyrtomium,Genus Diacalpe, Genus Didymochlaena, Genus Dryopsis, Genus Dryopteris,Genus Elaphoglossum, Genus Hypodematium, Genus Lastreopsis, GenusLeptorumohra, Genus Leucostegia, Genus Lithostegia, Genus Lomagramma,Genus Maxonia, Genus Megalastrum, Genus Olfersia, Genus Peranema, GenusPhanerophlebia, Genus Phanerophlebiopsis, Genus Polybotrya, GenusPolystichopsis, Genus Polystichum, Genus Rumohra, Genus Sorolepidium,Genus Stigmatopteris or Genus Teratophyllum. In some embodiments thenucleic acid molecule encoding the PtIP-50 polypeptide is derived from afern species in the Order Polypodiales, Family Dryopteridaceae, GenusBolbitis, selected from but not limited to Bolbitis acrostichoides,Bolbitis aliena, Bolbitis angustipinna, Bolbitis appendiculata, Bolbitisauriculata, Bolbitis bemoullii, Bolbitis bipinnatifida, Bolbitiscadieri, Bolbitis christensenfi, Bolbitis confertifolia, Bolbitiscostata, Bolbitis crispatula, Bolbitis fluviatilis, Bolbitisgaboonensis, Bolbitis gemmifera, Bolbitis hainanensis, Bolbitis hastata,Bolbitis hekouensis, Bolbitis hemiotis, Bolbitis heteroclita, Bolbitisheudelotii, Bolbitis humblotii, Bolbitis interlineata, Bolbitislatipinna, Bolbitis laxireticulata, Bolbitis lindigfi, Bolbitislonchophora, Bolbitis longiflagellata, Bolbitis major, Bolbitis media,Bolbitis nicotianifolia, Bolbitis nodiflora, Bolbitis novoguineensis,Bolbitis oligarchica, Bolbitis palustris, Bolbitis pandurifolia,Bolbitis pergamentacea, Bolbitis portoricensis, Bolbitis presliana,Bolbitis quoyana, Bolbitis rawsonfi, Bolbitis repanda, Bolbitisrhizophylla, Bolbitis riparia, Bolbitis rivularis, Bolbitis sagenioides,Bolbitis salicina, Bolbitis scalpturata, Bolbitis scandens, Bolbitissemicordata, Bolbitis semipinnatifida, Bolbitis serrata, Bolbitisserratifolia, Bolbitis simplex, Bolbitis sinensis, Bolbitissingaporensis, Bolbitis sinuata, Bolbitis subcordata, Bolbitissubcrenata, Bolbitis taylorii, Bolbitis tibetica, Bolbitis tonkinensis,Bolbitis umbrosa, Bolbitis vanuaensis, and Bolbitis virens.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Lomariopsidaceae, Genus Nephrolepis. In some embodiments thenucleic acid molecule encoding the PtIP-50 polypeptide is derived from afern species in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis is selected from but not limited to Nephrolepis abrupta,Nephrolepis acuminata, Nephrolepis acutifolia, Nephrolepis arida,Nephrolepis arthropteroides, Nephrolepis biserrata var. auriculata,Nephrolepis brownii, Nephrolepis celebica, Nephrolepis clementis,Nephrolepis cordifolia, Nephrolepis davalliae, Nephrolepis davallioides,Nephrolepis dayakorum, Nephrolepis delicatula, Nephrolepisdicksonioides, Nephrolepis duffii, Nephrolepis exaltata ssp. exaltatassp. Hawaiiensis, Nephrolepis falcata, Nephrolepis falciformis,Nephrolepis glabra, Nephrolepis hirsutula, Nephrolepis humatoides,Nephrolepis iridescens, Nephrolepis kurotawae, Nephrolepis laurifolia,Nephrolepis lauterbachii, Nephrolepis lindsayae, Nephrolepis multifida,Nephrolepis multiflora, Nephrolepis niphoboloides, Nephrolepisobliterate, Nephrolepis paludosa, Nephrolepis pectinata, Nephrolepispendula, Nephrolepis persicifolia, Nephrolepis pickelii, Nephrolepispilosula, Nephrolepis pubescens, Nephrolepis pumicicola, Nephrolepisradicans, Nephrolepis rivularis, Nephrolepis rosenstockii, Nephrolepissaligna, Nephrolepis schlechteri, Nephrolepis serrate, Nephrolepisthomsoni, Nephrolepis undulata var. aureoglandulosa, Nephrolepis×averyi,Nephrolepis×copelandii, and Nephrolepis×medlerae.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Polypodiaceae, Genus Campyloneurum, Genus Drynaria, GenusLepisorus, Genus Microgramma, Genus Microsorum, Genus Neurodium, GenusNiphidium, Genus Pecluma M.G., Genus Phlebodium, Genus Phymatosorus,Genus Platycerium, Genus Pleopeltis, Genus Polypodium L.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Polypodiaceae, Genus Polypodium L. In some embodiments thenucleic acid molecule encoding the PtIP-50 polypeptide is derived from afern species in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L. selected from but not limited to Polypodium absidatum,Polypodium acutifolium, Polypodium adiantiforme, Polypodium aequale,Polypodium affine, Polypodium albidopaleatum, Polypodium alcicorne,Polypodium alfarii, Polypodium alfredii, Polypodium alfredii var.curtii, Polypodium allosuroides, Polypodium alsophilicola, Polypodiumamamianum, Polypodium amoenum, Polypodium amorphum, Polypodiumanetioides, Polypodium anfractuosum, Polypodium anguinum, Polypodiumangustifolium f. remotifolia, Polypodium angustifolium var. amphostenon,Polypodium angustifolium var. heterolepis, Polypodium angustifolium var.monstrosa, Polypodium angustipaleatum, Polypodium angustissimum,Polypodium anisomeron var. pectinatum, Polypodium antioquianum,Polypodium aoristisorum, Polypodium apagolepis, Polypodium apicidens,Polypodium apiculatum, Polypodium apoense, Polypodium appalachianum,Polypodium appressum, Polypodium arenarium, Polypodium argentinum,Polypodium argutum, Polypodium armaturn, Polypodium aromaticum,Polypodium aspersum, Polypodium assurgens, Polypodium atrum, Polypodiumauriculatum, Polypodium balaonense, Polypodium balliviani, Polypodiumbamleri, Polypodium bangii, Polypodium bartlettii, Polypodium basale,Polypodium bemoullii, Polypodium biauritum, Polypodium bifrons,Polypodium blepharodes, Polypodium bolivari, Polypodium bolivianurn,Polypodium bolobense, Polypodium bombycinum, Polypodium bombycinum var.insularum, Polypodium bradeorum, Polypodium bryophilum, Polypodiumbryopodum, Polypodium buchtienii, Polypodium buesii, Polypodiumbulbotrichum, Polypodium caceresii, Polypodium californicum f.brauscombii, Polypodium californicum f. parsonsiae, Polypodiumcalifornicum, Polypodium calophlebium, Polypodium calvum, Polypodiumcamptophyllarium var. abbreviatum, Polypodium capitellatum, Polypodiumcarpinterae, Polypodium chachapoyense, Polypodium chartaceum, Polypodiumchimantense, Polypodium chiricanum, Polypodium choquetangense,Polypodium christensenii, Polypodium christii, Polypodium chrysotrichum,Polypodium ciliolepis, Polypodium cinerascens, Polypodium collinsii,Polypodium colysoides, Polypodium confluens, Polypodium conforme,Polypodium confusum, Polypodium congregatifolium, Polypodium connellii,Polypodium consimile var. bourgaeanum, Polypodium consimile var. minor,Polypodium conterminans, Polypodium contiguum, Polypodium cookii,Polypodium coriaceum, Polypodium coronans, Polypodium costaricense,Polypodium costatum, Polypodium crassifolium f. angustissimum,Polypodium crassifolium var. longipes, Polypodium crassulum, Polypodiumcraterisorum, Polypodium cryptum, Polypodium crystalloneuron, Polypodiumcucullaturn var. planum, Polypodium cuencanum, Polypodium cumingianum,Polypodium cupreolepis, Polypodium curranii, Polypodium curvans,Polypodium cyathicola, Polypodium cyathisorum, Polypodium cyclocolpon,Polypodium daguense, Polypodium damunense, Polypodium dareiformioides,Polypodium dasypleura, Polypodium decipiens, Polypodium decorum,Polypodium delicatulum, Polypodium deltoideum, Polypodium demeraranum,Polypodium denticulatum, Polypodium diaphanum, Polypodium dilataturn,Polypodium dispersum, Polypodium dissectum, Polypodium dissimulans,Polypodium dolichosorum, Polypodium dolorense, Polypodiumdonnell-smithii, Polypodium drymoglossoides, Polypodium ebeninum,Polypodium eggersii, Polypodium elmeri, Polypodium elongatum, Polypodiumenterosoroides, Polypodium erubescens, Polypodium erythrolepis,Polypodium erythrotrichum, Polypodium eurybasis, Polypodium eurybasisvar. villosum, Polypodium exornans, Polypodium falcoideum, Polypodiumfallacissimum, Polypodium farinosum, Polypodium faucium, Polypodiumfeel, Polypodium ferrugineum, Polypodium feuillei, Polypodium firmulum,Polypodium firmum, Polypodium flaccidum, Polypodium flagellare,Polypodium flexuosum, Polypodium flexuosum var. ekmanii, Polypodiumforbesii, Polypodium formosanum, Polypodium fraxinifolium subsp.articulatum, Polypodium fraxinifolium subsp. luridum, Polypodiumfructuosum, Polypodium fucoides, Polypodium fulvescens, Polypodiumgaleottii, Polypodium glaucum, Polypodium glycyrrhiza, Polypodiumgracillimum, Polypodium gramineum, Polypodium grandifolium, Polypodiumgratum, Polypodium graveolens, Polypodium griseo-nigrum, Polypodiumgriseum, Polypodium guttatum, Polypodium haalilioanum, Polypodiumhammatisorum, Polypodium hancockii, Polypodium haplophlebicum,Polypodium harrisii, Polypodium hastatum var. simplex, Polypodiumhawaiiense, Polypodium heanophyllum, Polypodium helleri, Polypodiumhemionitidium, Polypodium henryi, Polypodium herzogii, Polypodiumhesperium, Polypodium hessii, Polypodium hombersleyi, Polypodiumhostmannii, Polypodium humile, Polypodium hyalinum, Polypodium iboense,Polypodium induens var. subdentatum, Polypodium insidiosum, Polypodiuminsigne, Polypodium intermedium subsp. masafueranum var. obtuseserratum,Polypodium intramarginale, Polypodium involutum, Polypodium itatiayense,Polypodium javanicum, Polypodium juglandifolium, Polypodium kaniense,Polypodium knowltoniorum, Polypodium kyimbilense, Polypodiuml'herminieri var. costaricense, Polypodium lachniferum f. incurvata,Polypodium lachniferum var. glabrescens, Polypodium lachnopus,Polypodium lanceolatum var. complanatum, Polypodium lanceolatum var.trichophorum, Polypodium latevagans, Polypodium laxifrons, Polypodiumlaxifrons var. lividum, Polypodium lehmannianum, Polypodium leiorhizum,Polypodium leptopodon, Polypodium leuconeuron var. angustifolia,Polypodium leuconeuron var. latifolium, Polypodium leucosticta,Polypodium limulum, Polypodium lindigii, Polypodium lineatum, Polypodiumlomarioides, Polypodium longifrons, Polypodium loretense, Polypodiumloriceum var. umbraticum, Polypodium loriforme, Polypodium loxogramme f.gigas, Polypodium ludens, Polypodium luzonicum, Polypodium lycopodioidesf. obtusum, Polypodium lycopodioides L., Polypodium macrolepis,Polypodium macrophyllum, Polypodium macrosorum, Polypodiummacrosphaerum, Polypodium maculosum, Polypodium madrense, Polypodiummanmeiense, Polypodium margaritiferum, Polypodium maritimum, Polypodiummartensii, Polypodium mayoris, Polypodium megalolepis, Polypodiummelanotrichum, Polypodium menisciifolium var. pubescens, Polypodiummeniscioides, Polypodium merrillii, Polypodium mettenii, Polypodiummexiae, Polypodium microsorum, Polypodium militare, Polypodium minimum,Polypodium minusculum, Polypodium mixtum, Polypodium mollendense,Polypodium mollissimum, Polypodium moniliforme var. minus, Polypodiummonoides, Polypodium monticola, Polypodium montigenum, Polypodiummoritzianum, Polypodium moultonii, Polypodium multicaudatum, Polypodiummultilineatum, Polypodium multisorum, Polypodium munchii, Polypodiummuscoides, Polypodium myriolepis, Polypodium myriophyllum, Polypodiummyriotrichum, Polypodium nematorhizon, Polypodium nemorale, Polypodiumnesioticum, Polypodium nigrescentium, Polypodium nigripes, Polypodiumnigrocinctum, Polypodium nimbatum, Polypodium nitidissimum, Polypodiumnitidissimum var. latior, Polypodium nubrigenum, Polypodium oligolepis,Polypodium oligosorum, Polypodium oligosorum, Polypodium olivaceum,Polypodium olivaceum var. elatum, Polypodium oodes, Polypodiumoosphaerum, Polypodium oreophilum, Polypodium ornatissimum, Polypodiumornatum, Polypodium ovatum, Polypodium oxylobum, Polypodium oxypholis,Polypodium pakkaense, Polypodium pallidum, Polypodium palmatopedatum,Polypodium palmeri, Polypodium panamense, Polypodium parvum, Polypodiumpatagonicum, Polypodium paucisorum, Polypodium pavonianum, Polypodiumpectinatum var. caliense, Polypodium pectinatum var. hispidum,Polypodium pellucidum, Polypodium pendulum var. boliviense, Polypodiumpercrassum, Polypodium perpusillum, Polypodium peruvianum var.subgibbosum, Polypodium phyllitidis var. elongatum, Polypodiumpichinchense, Polypodium pilosissimum, Polypodium pilosissimum var.glabriusculum, Polypodium pilossimum var. tunguraquensis, Polypodiumpityrolepis, Polypodium platyphyllum, Polypodium playfairii, Polypodiumplebeium var. cooperi, Polypodium plectolepidioides, Polypodiumpleolepis, Polypodium plesiosorum var.i, Polypodium podobasis,Polypodium podocarpum, Polypodium poloense, Polypodium polydatylon,Polypodium polypodioides var. aciculare, Polypodium polypodioides var.michauxianum, Polypodium praetermissum, Polypodium preslianum var.immersum, Polypodium procerum, Polypodium procerum, Polypodiumproductum, Polypodium productum, Polypodium prolongilobum, Polypodiumpropinguum, Polypodium proteus, Polypodium pruinatum, Polypodiumpseudocapillare, Polypodium pseudofratemum, Polypodium pseudonutans,Polypodium pseudoserratum, Polypodium pulcherrimum, Polypodiumpulogense, Polypodium pungens, Polypodium purpusii, Polypodium radicale,Polypodium randallii, Polypodium ratiborii, Polypodium reclinaturn,Polypodium recreense, Polypodium repens var. abruptum, Polypodiumrevolvens, Polypodium rhachipterygium, Polypodium rhomboideum,Polypodium rigens, Polypodium robustum, Polypodium roraimense,Polypodium roraimense, Polypodium rosei, Polypodium rosenstockii,Polypodium rubidum, Polypodium rudimentum, Polypodium rusbyi, Polypodiumsablanianum, Polypodium sarmentosum, Polypodium saxicola, Polypodiumschenckii, Polypodium schlechteri, Polypodium scolopendria, Polypodiumscolopendria, Polypodium scolopendrium, Polypodium scouleri, Polypodiumscutulatum, Polypodium segregatum, Polypodium semihirsutum, Polypodiumsemihirsutum var. fuscosetosum, Polypodium senile var. minor, Polypodiumsericeolanatum, Polypodium serraeforme, Polypodium serricula, Polypodiumsesquipedala, Polypodium sessilifolium, Polypodium setosum var. calvum,Polypodium setulosum, Polypodium shaferi, Polypodium sibomense,Polypodium siccum, Polypodium simacense, Polypodium simulans, Polypodiumsingeri, Polypodium sinicum, Polypodium sintenisii, Polypodium skutchii,Polypodium sloanei, Polypodium sodiroi, Polypodium sordidulum,Polypodium sordidum, Polypodium sphaeropteroides, Polypodium sphenodes,Polypodium sprucei, Polypodium sprucei var. furcativenosa, Polypodiumsteirolepis, Polypodium stenobasis, Polypodium stenolepis, Polypodiumstenopterum, Polypodium subcapillare, Polypodium subflabelliforme,Polypodium subhemionitidium, Polypodium subinaequale, Polypodiumsubintegrum, Polypodium subspathulatum, Polypodium subtile, Polypodiumsubvestitum, Polypodium subviride, Polypodium superficiale var.attenuatum, Polypodium superficiale var. chinensis, Polypodiumsursumcurrens, Polypodium tablazianurn, Polypodium taenifolium,Polypodium tamandarei, Polypodium tatei, Polypodium tenuiculum var.acrosora, Polypodium tenuiculum var. brasiliense, Polypodium tenuilore,Polypodium tenuinerve, Polypodium tepuiense, Polypodium teresae,Polypodium tetragonum var. incompletum, Polypodium thysanolepis var.bipinnatifidum, Polypodium thyssanolepis, var. thyssanolepis, Polypodiumthyssanolepsi, Polypodium tobagense, Polypodium trichophyllum,Polypodium tridactylum, Polypodium tridentatum, Polypodium trifurcaturnvar. brevipes, Polypodium triglossum, Polypodium truncatulum, Polypodiumtruncicola var. major, Polypodium truncicola var. minor, Polypodiumtuberosum, Polypodium tunguraguae, Polypodium turquinurn, Polypodiumturrialbae, Polypodium ursipes, Polypodium vagans, Polypodiumvaldealaturn, Polypodium versteegii, Polypodium villagranii, Polypodiumvirginianum f. cambroideum, Polypodium virginianurn f. peraferens,Polypodium vittarioides, Polypodium vulgare, Polypodium vulgare L.,Polypodium vulgare subsp. oreophilum, Polypodium vulgare var.acuminatum, Polypodium vulpinum, Polypodium williamsii, Polypodiumwobbense, Polypodium×fallacissimum-guttatum, Polypodium xantholepis,Polypodium xiphopteris, Polypodium yarumalense, Polypodium yungense, andPolypodium zosteriforme.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Order Polypodiales,Family Polypodiaceae, Genus Platycerium. In some embodiments the nucleicacid molecule encoding the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium selected from but not limited to Platycerium alcicorne,Platycerium andinum, Platycerium angolense, Platycerium bifurcatum,Platycerium coronarium, Platycerium elephantotis, Platycerium ellisfi,Platycerium grande, Platycerium hillii, Platycerium holttumfi,Platycerium madagascariense, Platycerium quadridichotomum, Platyceriumridleyi, Platycerium sp. ES-2011, Platycerium stemaria, Platyceriumsuperbum, Platycerium veitchii, Platycerium wallichii, Platyceriumwandae, Platycerium wilhelminae-reginae, and Platycerium willinckii.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Division Lycophyta. Thephylogeny of extant Lycopods as used herein is based on theclassification by N. Wikstrom, American Fern Journal, 91:150-156 (2001).Other phylogenic classifications of extant Lycopods are known to oneskilled in the art. Additional information on the phylogeny of ferns canbe found at mobot.org/MOBOT/research/APweb/(which can be accessed usingthe “www” prefix) and Schuettpelz E. and Pryer K. M., TAXON 56:1037-1050 (2007) based on three plastid genes. Additional Lycopodspecies can be found at homepages.caverock.net.nz/˜byfern/list.htm(which can be accessed using the http:// prefix).

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Class Isoetopsida or ClassLycopodiopsida.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Class Isoetopsida OrderSelaginales. In some embodiments the nucleic acid molecule encoding thePtIP-50 polypeptide is derived from a fern species in the ClassIsoetopsida, Order Selaginales, Family Selaginellaceee. In someembodiments the nucleic acid molecule encoding the PtIP-50 polypeptideis derived from a species in the Genus Selaginella. In some embodimentsthe nucleic acid molecule encoding the PtIP-50 polypeptide is derivedfrom a Selaginella species selected from but not limited to Selaginellaacanthonota, Selaginella apoda, Selaginella arbuscula, Selaginellaarenicola, Selaginella arizonica, Selaginella armata, Selaginellaasprella, Selaginella biformis, Selaginella bigelovii, Selaginellabraunii, Selaginella cinerascens, Selaginella cordifolia, Selaginelladeflexa, Selaginella delicatula, Selaginella densa, Selaginelladouglasii, Selaginella eatonii, Selaginella eclipes, Selaginellaeremophila, Selaginella erythropus, Selaginella flabellata, Selaginellahansenii, Selaginella heterodonta, Selaginella kraussiana, Selaginellakrugii, Selaginella laxifolia, Selaginella lepidophylla, Selaginellaleucobryoides, Selaginella ludoviciana, Selaginella mutica, Selaginellaoregana, Selaginella ovifolia, Selaginella pallescens, Selaginellaperuviana, Selaginella pilifera, Selaginella plane, Selaginella plumose,Selaginella pulcherrima, Selaginella rupestris, Selaginella rupincola,Selaginella scopulorum, Selaginella selaginoides, Selaginella sibirica,Selaginella standleyi, Selaginella stellate, Selaginella subcaulescens,Selaginella substipitata, Selaginella tenella, Selaginella tortipila,Selaginella uliginosa, Selaginella umbrosa, Selaginella uncinata,Selaginella underwoodii, Selaginella utahensis, Selaginella victoriae,Selaginella viridissima, Selaginella wallacei, Selaginella watsonii,Selaginella weatherbiana, Selaginella willdenowii, Selaginella wrightiiand Selaginella×neomexicana.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Class Lycopodiopsida, OrderLycopodiales. In some embodiments the nucleic acid molecule encoding thePtIP-50 polypeptide is derived from a fern species in the ClassLycopodiopsida, Order Lycopodiales Family Lycopodiaceae or FamilyHuperziaceae. In some embodiments the nucleic acid molecule encoding thePtIP-50 polypeptide is derived from a species in the GenusAustrolycopodium, Dendrolycopodium, Diphasiastrum, Diphasium, Huperzia,Lateristachys, Lycopodiastrum, Lycopodiella, Lycopodium, Palhinhaea,Pseudodiphasium, Pseudolycopodiella, Pseudolycopodium or Spinulum. Insome embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Genus Lycopodium. In someembodiments the nucleic acid molecule encoding the PtIP-50 polypeptideis derived from a Lycopodium species selected from but not limited toLycopodium alpinum L., Lycopodium annotinum L., Lycopodium clavatum L.,Lycopodium cornplanatum L., Lycopodium dendroideum Michx., Lycopodiumdigitatum, Lycopodium xhabereri, Lycopodium hickeyi, Lycopodium×issleri,Lycopodium lagopus, Lycopodium obscurum L., Lycopodium phlegmaria L.,Lycopodium sabinifolium, Lycopodium sitchense, Lycopodium tristachyum,Lycopodium venustulum, Lycopodium venustulum var. venustulum, Lycopodiumvenustulum var. verticale, Lycopodium volubile and Lycopodium×zeilleri.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a species in the Genus Huperzia. In someembodiments the nucleic acid molecule encoding the PtIP-50 polypeptideis derived from a species selected from but not limited to Huperziaappressa, Huperzia arctica, Huperzia attenuata, Huperzia australiana,Huperzia balansae, Huperzia billardierei, Huperzia brassii, Huperziacampiana, Huperzia capellae, Huperzia carinata, Huperzia cf. carinataARF000603, Huperzia cf. nummulariifolia ARF001140, Huperzia cf.phlegmaria ARF000717, Huperzia cf. phlegmaria ARF000771, Huperzia cf.phlegmaria ARF000785, Huperzia cf. phlegmaria ARF001007, Huperzia cf.phlegmaria ARF002568, Huperzia cf. phlegmaria ARF002703, Huperzia cf.phlegmaria Wikstrom 1998, Huperzia chinensis, Huperzia compacta,Huperzia crassa, Huperzia crispata, Huperzia ctyptomeriana, Huperziacumingii, Huperzia dacrydioides, Huperzia dalhousieana, Huperziadichotoma, Huperzia emeiensis, Huperzia ericifolia, Huperzia eversa,Huperzia fargesii, Huperzia fordii, Huperzia funiformis, Huperziagoebellii, Huperzia haleakalae, Huperzia hamiltonii, Huperziaheteroclita, Huperzia hippuridea, Huperzia hippuris, Huperzia holstii,Huperzia horizontalis, Huperzia hunanensis, Huperzia hystrix, Huperzialindenii, Huperzia linifolia, Huperzia lockyeri, Huperzia lucidula,Huperzia mingcheensis, Huperzia miyoshiana, Huperzia nanchuanensis,Huperzia nummulariifolia, Huperzia obtusifolia, Huperziaophioglossoides, Huperzia petiolate, Huperzia phlegmaria, Huperziaphlegmarioides, Huperzia phyllantha, Huperzia pinifolia, Huperziapolydactyla, Huperzia prolifera, Huperzia reflexa, Huperziarosenstockiana, Huperzia rufescens, Huperzia salvinoides, Huperziasarmentosa, Huperzia selago, Huperzia serrata, Huperzia sieboldii,Huperzia somae, Huperzia squarrosa, Huperzia subulata, Huperziasutchueniana, Huperzia tauri, Huperzia taxifolia, Huperzia tenuis,Huperzia tetragona, Huperzia tetrasticha, Huperzia unguiculata, Huperziavaria, Huperzia verticillata and Huperzia wilsonii.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising at least one amino acid sequence motif selectedfrom: an amino acid sequence motif 1 as represented by an amino acidsequence of the formulaAWK[AT][KS]C[KR]NVA[AV][LV]G[RQ]E[ML][CTS]TTAA[YH]I[SA][EQ]LQY[DEQ]I[QK]VQ[AE]MLQ[EDQ]IA[RQ][KR]QA[DE]RL[SE][SGA]I(SEQ ID NO: 123); an amino acid sequence motif 2 as represented by anamino acid sequence of the formula[EQ][SA]YDQ[ESD][LF][KR]Q[LF][KQ]LF[IV][AQ]QN[KE]ILG[SG]YLL[EQ]QN[RK]AFA[EA][KR]E[RK]DM[ED][VA]FHS(SEQ ID NO: 124); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaF[HR]DRN[EK][TASGERDN]E[VP]L[HE]YEA[AG][TV][PG]L[DV]Y[HQ]YAY[NR]LDTG[EA]TT[LV][TS]NLPS(SEQ ID NO: 125); an amino acid sequence motif 4 as represented by anamino acid sequence of the formulaADL[ST][SN][YF]TE[ML][VA][TS][QE]MDMRTTR[LM]L[LV][EA]LIKVL[HNY][IM]QN[AGV]ALMY[QE]YLS[EP][PA] SEQ ID NO: 126); an amino acid sequence motif 5 asrepresented by an amino acid sequence of the formulaPFT[RNT]WRLR[LV]SASA[EQ]EN[EKQ]GLAFP (SEQ ID NO: 127); an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaDW[LD]IF[VE]NE[VI]E[AG]VA[EA]QMP[TE]EVSE (SEQ ID NO: 128); an amino acidsequence motif 7 as represented by an amino acid sequence of the formulaV[TN]M[EDQ]TVW[GR][ML]L[VI]Q[HQ]E[HQ]AA[IV][LQ]GL[LM][RQ]LG[PA][SAP][FS]D[FI][TRV]RT[YF]VVKDIPV[DS]LLL[DH]G (SEQ ID NO: 129); an amino acid sequencemotif 8 as represented by an amino acid sequence of the formulaIRLD[QK]MEFSEVM[VA][IV]HRM[FH][IVF][RK][LM][DP][DE]LD[IVL][AG][HL] (SEQID NO: 130); an amino acid sequence motif 9 as represented by an aminoacid sequence of the formula[AL]A[RE][GK]L[QKE][RK]V[LV][EQ]IL[ED][GD]LQA[VA]MEVV[AK][AI]I[KN][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][ML]P[TS]E (SEQ ID NO: 131); an amino acidsequence motif 10 as represented by an amino acid sequence of theformula [TV]FP[LS][TS][WL]SRVRI[HR][HY][LV]EMKF[VD][GQA][AG]A (SEQ IDNO: 132); an amino acid sequence motif 11 as represented by an aminoacid sequence of the formula[RK]Q[VL]A[RN]A[LV]FAV[LF][GR]AI[AG][AS][VI][AG]L[AT][FVL][ALV]TGGA (SEQID NO: 133); an amino acid sequence motif 12 as represented by an aminoacid sequence of the formula[QL][RK][QRT][STI]EL[DQ]N[TA][IML][QE][KR][MI]DQLSLQMET[QE]S[AE][DA]M[ED]QA[KQR][AE]DM[ED][AE] (SEQ ID NO: 134); an amino acid sequence motif 13 asrepresented by an amino acid sequence of the formulaLVVP[QR]LQY[DH]MYSNLI[ND][RQ][ML][AV][QR]VA (SEQ ID NO: 135); an aminoacid sequence motif 14 as represented by an amino acid sequence of theformula[PA][DV]A[LV]LTDP[NS][IV][LI][LS][GC]MQT[ST][ML]LIAELV[EL][FV][AS][HQ][PN]SS(SEQ ID NO: 136); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[AT][TV][SG]A[DG][AD]TTQIAITF[HYF][VI][ST]AIR[RQE]I (SEQ ID NO: 137); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula LEGA[ED]KVKRLYVFADVVEL[PE][SG]K (SEQ ID NO: 138); an aminoacid sequence motif 17 as represented by an amino acid sequence of theformula P[IV]TD[TS]GE[VI]Y[IM]LLQ[SG]SR (SEQ ID NO: 139); an amino acidsequence motif 18 as represented by an amino acid sequence of theformula T[VL][RS]LPG[ST][IV][MS]VVILCR[VI]L[HV][LV]N[GS] (SEQ ID NO:140); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula [AV]V[TR][KR]H[VA]EWLN[KT]LL[LV]Q[AV]SAAAQ[GS](SEQ ID NO: 141); an amino acid sequence motif 20 as represented by anamino acid sequence of the formula[YV][LV]ALL[FY]R[AT]Q[YA][LVF][IL]K[LGM]V[GK] (SEQ ID NO: 142); an aminoacid sequence motif 21 as represented by an amino acid sequence of theformula AP[GAL]A[VIM][AS]AA[KGR][GA]AV[ST][AI]AG (SEQ ID NO: 143); anamino acid sequence motif 22 as represented by an amino acid sequence ofthe formula [ED]D[WY][EQ]FE[IL]PVEDF (SEQ ID NO: 144); an amino acidsequence motif 23 as represented by an amino acid sequence of theformula YS[EH]LYR[DE][LV]NQ[VI]S (SEQ ID NO: 145); and an amino acidsequence motif 24 as represented by an amino acid sequence of theformula F[AI][NR]TFMRMT (SEQ ID NO: 146).

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising at least one amino acid sequence motif selectedfrom: an amino acid sequence motif 1 having at least 90% identity to anamino acid sequence of AWK[AT][KS]C[KR] NVA[AV][LV]G[RQ]E[ML][CTS]TTAA[YH]I[SA][EQ]LQY[DEQ]I[QK]VQ[AE]MLQ[EDQ]IA[RQ][KR]QA[DE]RL[SE][SGA]I (SEQ ID NO:123); an amino acid sequence motif 2 having at least 90% identity to anamino acid sequence of[EQ][SA]YDQ[ESD][LF][KR]Q[LF][KQ]LF[IV][AQ]QN[KE]ILG[SG]YLL[EQ]QN[RK]AFA[EA][KR]E[RK]DM[ED][VA]FHS(SEQ ID NO: 124); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence ofF[HR]DRN[EK][TASGERDN]E[VP]L[HE]YEA[AG][TV][PG]L[DV]Y[HQ]YAY[NR]LDTG[EA]TT[LV][TS]NLPS(SEQ ID NO: 125); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence ofADL[ST][SN][YF]TE[ML][VA][TS][QE]MDMRTTR[LM]L[LV][EA]LIKVL[HNY][IM]QN[AGV]ALMY[QE]YLS[EP][PA] (SEQ ID NO: 126); an amino acid sequence motif 5 havingat least 90% identity to an amino acid sequence ofPFT[RNT]WRLR[LV]SASA[EQ]EN[EKQ]GLAFP (SEQ ID NO: 127); an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof DW[LD]IF[VE]NE[VI]E[AG]VA[EA]QMP[TE]EVSE (SEQ ID NO: 128); an aminoacid sequence motif 7 having at least 90% identity to an amino acidsequence ofV[TN]M[EDQ]TVW[GR][ML]L[VI]Q[HQ]E[HQ]AA[IV][LQ]GL[LM][RQ]LG[PA][SAP][FS]D[FI][TRV]RT[YF]VVKDIPV[DS]LLL[DH]G (SEQ ID NO: 129); an amino acid sequencemotif 8 having at least 90% identity to an amino acid sequence ofIRLD[QK]MEFSEVM[VA][IV]HRM[FH][IVF][RK][LM][DP][DE]LD[IVL][AG][HL] (SEQID NO: 130); an amino acid sequence motif 9 having at least 90% identityto an amino acid sequence of[AL]A[RE][GK]L[QKE][RK]V[LV][EQ]IL[ED][GD]LQA[VA]MEVV[AK][AI]I[KN][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][ML]P[TS]E (SEQ ID NO: 131); an amino acidsequence motif 10 having at least 90% identity to an amino acid sequenceof [TV]FP[LS][TS][WL]SRVRI[HR][HY][LV]EMKF[VD][GQA][AG]A (SEQ ID NO:132); an amino acid sequence motif 11 having at least 90% identity to anamino acid sequence of[RK]Q[VL]A[RN]A[LV]FAV[LF][GR]AI[AG][AS][VI][AG]L[AT][FVL][ALV]TGGA (SEQID NO: 133); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of[QL][RK][QRT][STI]EL[DQ]N[TA][IML][QE][KR][MI]DQLSLQMET[QE]S[AE][DA]M[ED]QA[KQR][AE]DM[ED][AE] (SEQ ID NO: 134); an amino acid sequence motif 13having at least 90% identity to an amino acid sequence ofLVVP[QR]LQY[DH]MYSNLI[ND][RQ][ML][AV][QR]VA (SEQ ID NO: 135); an aminoacid sequence motif 14 having at least 90% identity to an amino acidsequence of[PA][DV]A[LV]LTDP[NS][IV][LI][LS][GC]MQT[ST][ML]LIAELV[EL][FV][AS][HQ][PN]SS(SEQ ID NO: 136); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of[AT][TV][SG]A[DG][AD]TTQIAITF[HYF][VI][ST]AIR[RQE]I (SEQ ID NO: 137); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of LEGA[ED]KVKRLYVFADVVEL[PE][SG]K (SEQ ID NO: 138); anamino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of P[IV]TD[TS]GE[VI]Y[IM]LLQ[SG]SR (SEQ ID NO: 139); anamino acid sequence motif 18 having at least 90% identity to an aminoacid sequence of T[VL][RS]LPG[ST][IV][MS]VVILCR[VI]L[HV][LV]N[GS] (SEQID NO: 140); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of[AV]V[TR][KR]H[VA]EWLN[KT]LL[LV]Q[AV]SAAAQ[GS] (SEQ ID NO: 141); anamino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of [YV][LV]ALL[FY]R[AT]Q[YA][LVF][IL]K[LGM]V[GK] (SEQ IDNO: 142); an amino acid sequence motif 21 having at least 90% identityto an amino acid sequence of AP[GAL]A[VIM][AS]AA[KGR][GA]AV[ST][AI]AG(SEQ ID NO: 143); an amino acid sequence motif 22 having at least 90%identity to an amino acid sequence of [ED]D[WY][EQ]FE[IL]PVEDF (SEQ IDNO: 144); an amino acid sequence motif 23 having at least 90% identityto an amino acid sequence of YS[EH]LYR[DE][LV]NQ[VI]S (SEQ ID NO: 145);an amino acid sequence motif 24 having at least 90% identity to an aminoacid sequence of F[AI][NR]TFMRMT (SEQ ID NO: 146).

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising at least one amino acid sequence motif selectedfrom: an amino acid sequence motif 1 as represented by an amino acidsequence of the formulaAWK[ATS][KSRT]C[KR]NVA[AVLI][LVI]G[RQKN]E[MLIV][CTS]TTAA[YHWF]I[SAT][EQDN]LQY[DEQN]I[QKNR]VQ[AED]MLQ[EDQN]IA[RQKN][KR]QA[DE]RL[SETD][SGAT]I (SEQ IDNO: 99); an amino acid sequence motif 2 as represented by an amino acidsequence of the formula[EQND][SAT]YDQ[ESDT][LFIV][KR]Q[LFIV][KQRN]LF[IVL][AQN]QN[KERD]ILG[SGT]YLL[EQDN]QN[RK]AFA[EAD][KR]E[RK]DM[ED][VAIL]FHS (SEQ ID NO: 100); an aminoacid sequence motif 3 as represented by an amino acid sequence of theformulaF[HRK]DRN[EKDR]XTASGERDNE[VPIL]L[HERKD]YEA[AG][TVSIL][PG]L[DVELI]Y[HQN]YAY[NRQK]LDTG[EAD]TT[LVI][TS]NLPS (SEQ ID NO: 101); an amino acid sequencemotif 4 as represented by an amino acid sequence of the formulaADL[ST][SNTQ][YFW]TE[MLIV][VAIL][TS][QEDN]MDMRTTR[LMIV]L[LVI][EAD]LIKVL[HNYWF][IMVL]QN[AGVIL]ALMY[QEDN]YLS[EPD][PA] SEQ ID NO: 102); an amino acidsequence motif 5 as represented by an amino acid sequence of the formulaPFT[RNTKQS]WRLR[LVI]SASA[EQDN]EN[EKQDRN]GLAFP (SEQ ID NO: 103); an aminoacid sequence motif 6 as represented by an amino acid sequence of theformula DW[LDIVE]IF[VEILD]NE[VIL]E[AG]VA[EAD]QMP[TESD]EVSE (SEQ ID NO:104); an amino acid sequence motif 7 as represented by an amino acidsequence of the formulaV[TNSQ]M[EDQN]TVW[GRK][MLIV]L[VIL]Q[HQN]E[HQN]AA[IVL][LQIVN]GL[LMIV][RQKN]LG[PA][SAPT][FSWYT]D[FILV][TRVSKIL]RT[YFW]VVKDIPV[DSET]LLL[DHE]G (SEQ IDNO: 105); an amino acid sequence motif 8 as represented by an amino acidsequence of the formulaIRLD[QKNR]MEFSEVM[VAIL][IVL]HRM[FHYW][IVFL][RK][LMIV][DPE][DE]LD[IVL][AG][HLIV](SEQ ID NO: 106); an amino acid sequence motif 9 as represented by anamino acid sequence of the formula[ALIV]A[REKD][GKR]L[QKENRD][RK]V[LVI][EQDN]IL[ED][GDE]LQA[VAIL]MEVV[AKR][AILV]I[KNRQ][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][MLIV]P[TS]E(SEQ ID NO: 107); an amino acid sequence motif 10 as represented by anamino acid sequence of the formula[TVSIL][FP[LSIVT][TS][WLYFVI]SRVRI[HRK][HYWF][LVI]EMKF[VDILE][GQAN][AG]A(SEQ ID NO: 108); an amino acid sequence motif 11 as represented by anamino acid sequence of the formula[RK]Q[VLI]A[RN]A[LVI]FAV[LFIV][GRK]AI[AG][AST][VIL][AG]L[ATS][FVLI][ALVI]TGGA(SEQ ID NO: 109); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[QLNIV][RK][QRTNKS][STILV]EL[DQEN]N[TAS][IMLV][QEDN][KR][MIVL]DQLSLQMET[QEDN]S[AED][DAE]M[ED]QA[KQRN][AED]DM[ED][AED] (SEQ ID NO: 110); an aminoacid sequence motif 13 as represented by an amino acid sequence of theformula LWP[QRNK]LQY[DHE]MYSNLI[NDQE][RQKN][MLIV][AVIL][QRNK]VA (SEQ IDNO: 111); an amino acid sequence motif 14 as represented by an aminoacid sequence of the formula[PA][DVELI]A[LVI]LTDP[NSQT][IVL][LIV][LSIVT][GC]MQT[ST][MLIV]LIAELV[ELDIV][FVIL][AST][HQN][PNQ]SS (SEQ ID NO: 112); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[ATS][TVSIL][SGT]A[DGE][ADE]TTQIAITF[HYFW][VIL][ST]AIR[RQEKND]I (SEQ IDNO: 113); an amino acid sequence motif 16 as represented by an aminoacid sequence of the formula LEGA[ED]KVKRLYVFADVVEL[PED][SGT]K (SEQ IDNO: 114); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula P[IVL]TD[TS]GE[VIL]Y[IMVL]LLQ[SGT]SR (SEQID NO: 115); an amino acid sequence motif 18 as represented by an aminoacid sequence of the formulaT[VLI][RSKT]LPG[ST][IVL][MST]WILCR[VIL]L[HVIL][LVI]N[GST] (SEQ ID NO:116); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula[AVIL]V[TR][KR]H[VAIL]EWLN[KT]LL[LVI]Q[AVIL]SAAAQ[GST] (SEQ ID NO: 117);an amino acid sequence motif 20 as represented by an amino acid sequenceof the formula [YV][LVI]ALL[FYW]R[ATS]Q[YAWF][LVFI][ILV]K[LGMIV]V[GKR](SEQ ID NO: 118); and an amino acid sequence motif 21 as represented byan amino acid sequence of the formulaAP[GALIV]A[VIML][AST]AA[KGR][GA]AV[ST][AIL]AG (SEQ ID NO: 119); an aminoacid sequence motif 22 as represented by an amino acid sequence of theformula [ED]D[WYF][EQDN]FE[ILV]PVEDF (SEQ ID NO: 120); an amino acidsequence motif 23 as represented by an amino acid sequence of theformula YS[EHD]LYR[DE][LVI]NQ[VIL]S (SEQ ID NO: 121) an amino acidsequence motif 24 as represented by an amino acid sequence of theformula F[AIV][NRQK]TFMRMT (SEQ ID NO: 122).

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising at least one amino acid sequence motif selectedfrom: an amino acid sequence motif 1 having at least 90% identity to anamino acid sequence ofAWK[ATS][KS]C[KR]NVA[AVIL][LVI]G[RQKN]E[MLIV][CTS]TTAA[YHWF]I[SAT][EQDN]LQY[DEQN]I[QKNR]VQ[AED]MLQ[EDQN]IA[RQKN][KR]QA[DE]RL[SETD][SGAT]I(SEQ ID NO: 99); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of[EQDN][SAT]YDQ[ESDT][LFIV][KR]Q[LFIV][KQRN]LF[IVL][AQN]QN[KERD]ILG[SGT]YLL[EQDN]QN[RK]AFA[EAD][KR]E[RK]DM[ED][VAIL]FHS (SEQ ID NO: 100); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence ofF[HRK]DRN[EKDR][TASGERDNKQ]E[VPIL]L[HERKD]YEA[AG][TVSIL][PG]L[DVELI]Y[HQN]YAY[NRQK]LDTG[EAD]TT[LVI][TS]NLPS(SEQ ID NO: 101); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence ofADL[ST][SNTQ][YFW]TE[MLIV][VAIL][TS][QEDN]MDMRTTR[LMIV]L[LVI][EAD]LIKVL[HNYWF][IMVL]QN[AGVIL]ALMY[QEDN]YLS[EPD][PA] (SEQ ID NO: 102); an amino acidsequence motif 5 having at least 90% identity to an amino acid sequenceof PFT[RNTKQS]WRLR[LVI]SASA[EQDN]EN[EKQDRN]GLAFP (SEQ ID NO: 103); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of DW[LDIVE]IF[VEILD]NE[VIL]E[AG]VA[EAD]QMP[TESD]EVSE (SEQID NO: 104); an amino acid sequence motif 7 having at least 90% identityto an amino acid sequence ofV[TNSQ]M[EDQN]TVW[GRK][MLIV]L[VIL]Q[HQN]E[HQN]AA[IVL][LQIVN]GL[LMIV][RQKN]LG[PA][SAPT][FSWYT]D[FILV][TRVSKIL]RT[YFW]VVKDIPV[DSET]LLL[DHE]G (SEQ IDNO: 105); an amino acid sequence motif 8 having at least 90% identity toan amino acid sequence ofIRLD[QKNR]MEFSEVM[VAIL][IVL]HRM[FHYW][IVFL][RK][LMIV][DPE][DE]LD[IVL][AG][HLIV](SEQ ID NO: 106); an amino acid sequence motif 9 having at least 90%identity to an amino acid sequence of[ALIV]A[REKD][GKR]L[QKENRD][RK]V[LVI][EQDN]IL[ED][GDE]LQA[VAIL]MEVV[AKR][AILV]I[KNRQ][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][MLIV]P[TS]E(SEQ ID NO: 107); an amino acid sequence motif 10 having at least 90%identity to an amino acid sequence of[TVSIL][FP[LSIVT][TS][WLYFVI]SRVRI[HRK][HYWF][LVI]EMKF[VDILE][GQAN][AG]A(SEQ ID NO: 108); an amino acid sequence motif 11 having at least 90%identity to an amino acid sequence of[RK]Q[VLI]A[RN]A[LVI]FAV[LFIV][GRK]AI[AG][AST][VIL][AG]L[ATS][FVLI][ALVI]TGGA(SEQ ID NO: 109); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of[QLNIV][RK][QRTNKS][STILV]EL[DQEN]N[TAS][IMLV][QEDN][KR][MIVL]DQLSLQMET[QEDN]S[AED][DAE]M[ED]QA[KQRN][AED]DM[ED][AED] (SEQ ID NO: 110); an aminoacid sequence motif 13 having at least 90% identity to an amino acidsequence of LWP[QRNK]LQY[DHE]MYSNLI[NDQE][RQKN][MLIV][AVIL][QRNK]VA (SEQID NO: 111); an amino acid sequence motif 14 having at least 90%identity to an amino acid sequence of[PA][DVELI]A[LVI]LTDP[NSQT][IVL][LIV][LSIVT][GC]MQT[ST][MLIV]LIAELV[ELDIV][FVIL][AST][HQN][PNQ]SS (SEQ ID NO: 112); an amino acid sequence motif 15 havingat least 90% identity to an amino acid sequence of[ATS][TVSIL][SGT]A[DGE][ADE]TTQIAITF[HYFW][VIL][ST]AIR[RQEKND]I (SEQ IDNO: 113); an amino acid sequence motif 16 having at least 90% identityto an amino acid sequence of LEGA[ED]KVKRLYVFADVVEL[PED][SGT]K (SEQ IDNO: 114); an amino acid sequence motif 17 having at least 90% identityto an amino acid sequence of P[IVL]TD[TS]GE[VIL]Y[IMVL]LLQ[SGT]SR (SEQID NO: 115); an amino acid sequence motif 18 having at least 90%identity to an amino acid sequence ofT[VLI][RSKT]LPG[ST][IVL][MST]WILCR[VIL]L[HVIL][LVI]N[GST] (SEQ ID NO:116); an amino acid sequence motif 19 having at least 90% identity to anamino acid sequence of[AVIL]V[TR][KR]H[VAIL]EWLN[KT]LL[LVI]Q[AVIL]SAAAQ[GST] (SEQ ID NO: 117);an amino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of [YV][LVI]ALL[FYW]R[ATS]Q[YAWF][LVFI][ILV]K[LGMIV]V[GKR](SEQ ID NO: 118); and an amino acid sequence motif 21 having at least90% identity to an amino acid sequence ofAP[GALIV]A[VIML][AST]AA[KGR][GA]AV[ST][AIL]AG (SEQ ID NO: 119); an aminoacid sequence motif 22 having at least 90% identity to an amino acidsequence of [ED]D[WYF][EQDN]FE[ILV]PVEDF (SEQ ID NO: 120); an amino acidsequence motif 23 having at least 90% identity to an amino acid sequenceof YS[EHD]LYR[DE][LVI]NQ[VIL]S (SEQ ID NO: 121), and an amino acidsequence motif 24 having at least 90% identity to an amino acid sequenceof F[AIV][NRQK]TFMRMT (SEQ ID NO: 122).

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising, sequentially from the N-terminus to theC-terminus, at least one amino acid sequence motif selected from: motif23; motif 8; motif 16; motif 18; motif 14; motif 19; motif 20; motif 13;motif 2; motif 12; motif 11; motif 21; motif 9; motif 6; motif 1; motif4; motif 7; motif 22; motif 10; motif 17; motif 3; motif 24; motif 5;and motif 15.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising, sequentially from the N-terminus to theC-terminus, the amino acid sequence motifs: motif 23; motif 8; motif 16;motif 18; motif 14; motif 19; motif 20; motif 13; motif 2; motif 12;motif 11; motif 21; motif 9; motif 6; motif 1; motif 4; motif 7; motif22; motif 10; motif 17; motif 3; motif 24; motif 5; and motif 15.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure; a RegionB of between about 380 to about 465 amino acids in length having aconsensus secondary structure comprising 8 to 10 segments ofpredominately alpha helical structure; and a Region C of between about150 to about 180 amino acids in length having a consensus secondarystructure comprising 6 to 8 segments of predominately beta strandstructure. As used herein “predominantly nonconserved secondarystructure” means that the regions of secondary structure don'tconsistently align within the family of PtIP-50 polypeptides. As usedherein “predominately alpha helical structure” means that secondarystructure prediction may have one or more gap of between 1 to 6 aminoacids of coil and/or beta strand structure intervening in the alphahelix structure. As used herein “predominately beta strand structure”means that secondary structure prediction may have one or more gap ofbetween 1 to 6 amino acids of coil and/or alpha helix structureintervening in the beta strand structure. In some embodiments thesecondary structure is generated by the PSIPRED, top ranked secondarystructure prediction method (Jones D T. (1999) J. Mol. Biol. 292:195-202).

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure; a RegionB of between about 380 to about 465 amino acids in length having aconsensus secondary structure comprising nine segments of predominatelyalpha helical structure; and a Region C of between about 150 to about180 amino acids in length having a consensus secondary structurecomprising seven segments of predominately beta strand structure.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure, whereinthe Region A comprises a conserved beta strand 1 (β1a) of between about4 and about 12 amino acids in length within about amino acid residue 50to about amino acid residue 130 from the N-terminus of the PtIP-50polypeptide; a Region B of between about 380 to about 465 amino acids inlength having a consensus secondary structure comprising nine segmentsof predominately alpha helical structure; and a Region C of betweenabout 150 to about 180 amino acids in length having a consensussecondary structure comprising seven segments of predominately betastrand structure.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a flexible consensus secondary structure, wherein theRegion A comprises a conserved beta strand 1 (β1a) of between about 4and about 12 amino acids in length, a coil of between about 3 and about18 amino acids in length and a beta strand 2 (β1b) of between about 4and about 32 amino acids in length, within about amino acid residue 50to about amino acid residue 165 from the N-terminus of the PtIP-50polypeptide; a Region B of between about 380 to about 465 amino acids inlength having a consensus secondary structure comprising nine segmentsof predominately alpha helical structure; and a Region C of betweenabout 150 to about 180 amino acids in length having a consensussecondary structure comprising seven segments of predominately betastrand structure.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure; a RegionB of between about 380 to about 465 amino acids in length having aconsensus secondary structure sequentially comprising i) an alphahelix-1 of between about 10 and about 26 amino acids in length; ii) acoil-1 of between about 2 and about 8 amino acids in length flanked byalpha helix-1 and alpha helix-2; iii) an alpha helix-2 of between about15 and about 24 amino acids in length; iv) a coil-2 of between about 4and about 14 amino acids in length flanked by alpha helix-2 and alphahelix-3; v) an alpha helix 3 of between about 10 and about 17 aminoacids in length; vi) a coil-3 of between about 11 and about 17 aminoacids in length flanked by alpha helix-3 and alpha helix-4; vii) analpha helix-4 of between about 24 and about 30 amino acids in length;viii) a coil-4 of between about 4 and about 13 amino acids in lengthflanked by alpha helix-4 and alpha helix-5; ix) an alpha helix-5 ofbetween about 48 and about 58 amino acids in length; x) a coil-5 ofbetween about 10 and about 15 amino acids in length flanked by alphahelix-5 and alpha helix-6; xi) an alpha helix-6 of between about 13 andabout 15 amino acids in length; xii) a coil-6 of between about 3 andabout 10 amino acids in length flanked by alpha helix-6 and alphahelix-7; xiii) an alpha helix-7 of between about 49 and about 59 aminoacids in length; xiv) a coil-7 of between about 0 and about 9 aminoacids in length flanked by alpha helix-7 and alpha helix-8; xv) an alphahelix-8 of between about 29 and about 36 amino acids in length; xvi) acoil-8 of between about 11 and about 16 amino acids in length flanked byalpha helix-8 and alpha helix-9; xvii) an alpha helix-9 of between about16 and about 22 amino acids in length; xviii) a coil-9 of between about5 and about 9 amino acids in length flanked by alpha helix-9 and RegionC; and a Region C of between about 150 to about 180 amino acids inlength having a consensus secondary structure comprising seven segmentsof predominately beta strand structure.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure; a RegionB of between about 380 to about 465 amino acids in length having aconsensus secondary structure comprising nine segments of predominatelyalpha helical structure; and a Region C of between about 150 to about180 amino acids in length having a consensus secondary structuresequentially comprising i) a beta strand-1 (β1) of between about 5 aminoacids and about 7 amino acids in length; ii) a coil of between about 12amino acids and about 17 amino acids in length; iii) a beta strand-2(β2) of between about 11 amino acids and about 15 amino acids in length;iv) a coil of between about 7 amino acids and about 23 amino acids inlength; v) a beta strand-3 (β3) of between about 5 amino acids and about8 amino acids in length; vi) a coil of between about 13 amino acids andabout 23 amino acids in length; vii) a beta strand-4 (β4) of betweenabout 3 amino acids and about 7 amino acids in length; viii) a coil ofbetween about 3 amino acids and about 8 amino acids in length; ix) abeta strand-5 (β5) of between about 4 amino acids and about 10 aminoacids in length; x) a coil of between about 24 amino acids and about 28amino acids in length; xi) a beta strand-6 (β6) of between about 5 aminoacids and about 7 amino acids in length; xii) a coil of between about 17amino acids and about 23 amino acids in length; and xiii) a betastrand-1 (β7) of between about 5 amino acids and about 7 amino acids inlength.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a predominantly nonconserved secondary structure; a RegionB of between about 380 to about 465 amino acids in length having aconsensus secondary structure sequentially comprising i) an alphahelix-1 of between about 10 and about 26 amino acids in length; ii) acoil-1 of between about 2 and about 8 amino acids in length flanked byalpha helix-1 and alpha helix-2; iii) an alpha helix-2 of between about15 and about 24 amino acids in length; iv) a coil-2 of between about 4and about 14 amino acids in length flanked by alpha helix-2 and alphahelix-3; v) an alpha helix 3 of between about 10 and about 17 aminoacids in length; vi) a coil-3 of between about 11 and about 17 aminoacids in length flanked by alpha helix-3 and alpha helix-4; vii) analpha helix-4 of between about 24 and about 30 amino acids in length;viii) a coil-4 of between about 4 and about 13 amino acids in lengthflanked by alpha helix-4 and alpha helix-5; ix) an alpha helix-5 ofbetween about 48 and about 58 amino acids in length; x) a coil-5 ofbetween about 10 and about 15 amino acids in length flanked by alphahelix-5 and alpha helix-6; xi) an alpha helix-6 of between about 13 andabout 15 amino acids in length; xii) a coil-6 of between about 3 andabout 10 amino acids in length flanked by alpha helix-6 and alphahelix-7; xiii) an alpha helix-7 of between about 49 and about 59 aminoacids in length; xiv) a coil-7 of between about 0 and about 9 aminoacids in length flanked by alpha helix-7 and alpha helix-8; xv) an alphahelix-8 of between about 29 and about 36 amino acids in length; xvi) acoil-8 of between about 11 and about 16 amino acids in length flanked byalpha helix-8 and alpha helix-9; xvii) an alpha helix-9 of between about16 and about 22 amino acids in length; xviii) a coil-9 of between about5 and about 9 amino acids in length flanked by alpha helix-9 and betastrand-1 (β1) of Region C; and a Region C of between about 150 to about180 amino acids in length having a consensus secondary structuresequentially comprising i) a beta strand-1 (β1) of between about 5 aminoacids and about 7 amino acids in length; ii) a coil of between about 12amino acids and about 17 amino acids in length; iii) a beta strand-2(β2) of between about 11 amino acids and about 15 amino acids in length;iv) a coil of between about 7 amino acids and about 23 amino acids inlength; v) a beta strand-3 (β3) of between about 5 amino acids and about8 amino acids in length; vi) a coil of between about 13 amino acids andabout 23 amino acids in length; vii) a beta strand-4 (β4) of betweenabout 3 amino acids and about 7 amino acids in length; viii) a coil ofbetween about 3 amino acids and about 8 amino acids in length; ix) abeta strand-5 (β5) of between about 4 amino acids and about 10 aminoacids in length; x) a coil of between about 24 amino acids and about 28amino acids in length; xi) a beta strand-6 (β6) of between about 5 aminoacids and about 7 amino acids in length; xii) a coil of between about 17amino acids and about 23 amino acids in length; and xiii) a betastrand-1 (β7) of between about 5 amino acids and about 7 amino acids inlength.

In some embodiments the nucleic acid molecule encodes a PtIP-50polypeptide comprising sequentially from the N-terminus to theC-terminus: a Region A of between about 200 to about 300 amino acids inlength having a flexible consensus secondary structure, wherein theRegion A comprises a conserved beta strand 1 (β1a) of between about 4and about 12 amino acids in length within about amino acid residue 50 toabout amino acid residue 130 from the N-terminus of the PtIP-50polypeptide; a Region B of between about 380 to about 465 amino acids inlength having a consensus secondary structure sequentially comprising i)an alpha helix-1 of between about 10 and about 26 amino acids in length;ii) a coil-1 of between about 2 and about 8 amino acids in lengthflanked by alpha helix-1 and alpha helix-2; iii) an alpha helix-2 ofbetween about 15 and about 24 amino acids in length; iv) a coil-2 ofbetween about 4 and about 14 amino acids in length flanked by alphahelix-2 and alpha helix-3; v) an alpha helix 3 of between about 10 andabout 17 amino acids in length; vi) a coil-3 of between about 11 andabout 17 amino acids in length flanked by alpha helix-3 and alphahelix-4; vii) an alpha helix-4 of between about 24 and about 30 aminoacids in length; viii) a coil-4 of between about 4 and about 13 aminoacids in length flanked by alpha helix-4 and alpha helix-5; ix) an alphahelix-5 of between about 48 and about 58 amino acids in length; x) acoil-5 of between about 10 and about 15 amino acids in length flanked byalpha helix-5 and alpha helix-6; xi) an alpha helix-6 of between about13 and about 15 amino acids in length; xii) a coil-6 of between about 3and about 10 amino acids in length flanked by alpha helix-6 and alphahelix-7; xiii) an alpha helix-7 of between about 49 and about 59 aminoacids in length; xiv) a coil-7 of between about 0 and about 9 aminoacids in length flanked by alpha helix-7 and alpha helix-8; xv) an alphahelix-8 of between about 29 and about 36 amino acids in length; xvi) acoil-8 of between about 11 and about 16 amino acids in length flanked byalpha helix-8 and alpha helix-9; xvii) an alpha helix-9 of between about16 and about 22 amino acids in length; xviii) a coil-9 of between about5 and about 9 amino acids in length flanked by alpha helix-9 and betastrand-1 (β1) of Region C; and a Region C of between about 150 to about180 amino acids in length having a consensus secondary structuresequentially comprising i) a beta strand-1 (β1) of between about 5 aminoacids and about 7 amino acids in length; ii) a coil of between about 12amino acids and about 17 amino acids in length; iii) a beta strand-2(β2) of between about 11 amino acids and about 15 amino acids in length;iv) a coil of between about 7 amino acids and about 23 amino acids inlength; v) a beta strand-3 (β3) of between about 5 amino acids and about8 amino acids in length; vi) a coil of between about 13 amino acids andabout 23 amino acids in length; vii) a beta strand-4 (β4) of betweenabout 3 amino acids and about 7 amino acids in length; viii) a coil ofbetween about 3 amino acids and about 8 amino acids in length; ix) abeta strand-5 (β5) of between about 4 amino acids and about 10 aminoacids in length; x) a coil of between about 24 amino acids and about 28amino acids in length; xi) a beta strand-6 (β6) of between about 5 aminoacids and about 7 amino acids in length; xii) a coil of between about 17amino acids and about 23 amino acids in length; and xiii) a betastrand-1 (β7) of between about 5 amino acids and about 7 amino acids inlength.

Polynucleotides Encoding PtIP-65 Polypeptides

One source of polynucleotides that encode PtIP-65 polypeptides orrelated proteins is a fern or other primitive plant species whichcontains a PtIP-65 polynucleotide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ IDNO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 or SEQ ID NO: 21 encoding aPtIP-65 polypeptide SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ IDNO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34,SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO:39, SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively. Thepolynucleotides of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20 and SEQ ID NO: 21 can be used to express PtIP-65polypeptides in bacterial hosts that include but are not limited toAgrobacterium, Bacillus, Escherichia, Salmonella, Pseudomonas andRhizobium bacterial host cells. The polynucleotides are also useful asprobes for isolating homologous or substantially homologouspolynucleotides that encode PtIP-65 polypeptides or related proteins.Such probes can be used to identify homologous or substantiallyhomologous polynucleotides derived from Pteridophyta species.

Polynucleotides that encode PtIP-65 polypeptides can also be synthesizedde novo from a PtIP-65 polypeptide sequence. The sequence of thepolynucleotide gene can be deduced from a PtIP-65 polypeptide sequencethrough use of the genetic code. Computer programs such as“BackTranslate” (GCG™ Package, Acclerys, Inc. San Diego, Calif.) can beused to convert a peptide sequence to the corresponding nucleotidesequence encoding the peptide. Examples of PtIP-65 polypeptidessequences that can be used to obtain corresponding nucleotide encodingsequences include, but are not limited to, the PtIP-65 polypeptide ofSEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ IDNO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40,SEQ ID NO: 41 and SEQ ID NO: 42. Furthermore, synthetic PtIP-65polynucleotide sequences of the disclosure can be designed so that theywill be expressed in plants. U.S. Pat. No. 5,500,365 describes a methodfor synthesizing plant genes to improve the expression level of theprotein encoded by the synthesized gene. This method relates to themodification of the structural gene sequences of the exogenoustransgene, to cause them to be more efficiently transcribed, processed,translated and expressed by the plant. Features of genes that areexpressed well in plants include elimination of sequences that can causeundesired intron splicing or polyadenylation in the coding region of agene transcript while retaining substantially the amino acid sequence ofthe toxic portion of the insecticidal protein. A similar method forobtaining enhanced expression of transgenes in monocotyledonous plantsis disclosed in U.S. Pat. No. 5,689,052.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is a polynucleotide having the sequence set forth in SEQ IDNO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ IDNO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20and SEQ ID NO: 21, and variants, fragments and complements thereof.“Complement” is used herein to refer to a nucleic acid sequence that issufficiently complementary to a given nucleic acid sequence such that itcan hybridize to the given nucleic acid sequence to thereby form astable duplex. “Polynucleotide sequence variants” is used herein torefer to a nucleic acid sequence that except for the degeneracy of thegenetic code encodes the same polypeptide.

In some embodiments the nucleic acid molecule encoding the PtIP-65polypeptide is a non-genomic nucleic acid sequence. As used herein a“non-genomic nucleic acid sequence” or “non-genomic nucleic acidmolecule” or “non-genomic polynucleotide” refers to a nucleic acidmolecule that has one or more change in the nucleic acid sequencecompared to a native or genomic nucleic acid sequence. In someembodiments the change to a native or genomic nucleic acid moleculeincludes but is not limited to: changes in the nucleic acid sequence dueto the degeneracy of the genetic code; codon optimization of the nucleicacid sequence for expression in plants; changes in the nucleic acidsequence to introduce at least one amino acid substitution, insertion,deletion and/or addition compared to the native or genomic sequence;removal of one or more intron associated with the genomic nucleic acidsequence; insertion of one or more heterologous introns; deletion of oneor more upstream or downstream regulatory regions associated with thegenomic nucleic acid sequence; insertion of one or more heterologousupstream or downstream regulatory regions; deletion of the 5′ and/or 3′untranslated region associated with the genomic nucleic acid sequence;insertion of a heterologous 5′ and/or 3′ untranslated region; andmodification of a polyadenylation site. In some embodiments thenon-genomic nucleic acid molecule is a cDNA. In some embodiments thenon-genomic nucleic acid molecule is a synthetic nucleic acid sequence.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is a non-genomic polynucleotide having a nucleotide sequencehaving at least 40%. 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%,59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%,73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%identity, to the nucleic acid sequence of SEQ ID NO: 1, SEQ ID NO: 2,SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7,SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12,SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 or SEQ ID NO: 21,wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33,SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO:38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42,wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO: 24, SEQ IDNO: 26 or SEQ ID NO: 29, wherein the PtIP-65 polypeptide in combinationwith a PtIP-50 polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 25, SEQ ID NO: 27, SEQ IDNO: 30, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO: 36,wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 23, SEQ ID NO: 28, SEQ IDNO: 31, SEQ ID NO: 32 or SEQ ID NO: 33, wherein the PtIP-65 polypeptidein combination with a PtIP-50 polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 41 or SEQ ID NO: 42,wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 38 or SEQ ID NO: 40,wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 37, wherein the PtIP-65polypeptide in combination with a PtIP-50 polypeptide has insecticidalactivity.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence having at least 40%, 45%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acidsequence of PtIP-65 polypeptide of SEQ ID NO: 39, wherein the PtIP-65polypeptide in combination with a PtIP-50 polypeptide has insecticidalactivity.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Division Pteridophyta.The phylogeny of ferns as used herein is based on the classification forextant ferns by A. R. Smith et al, TAXON, 55:705-731 (2006). Theconsensus phylogeny based on the classification by A. R. Smith is shownin FIG. 1. Additional information on the phylogeny of ferns can be foundat mobot.org/MOBOT/research/APweb/(which can be accessed using the “www”prefix) and Schuettpelz E. and Pryer K. M., TAXON 56: 1037-1050 (2007)based on three plastid genes. Additional fern and other primitive plantspecies can be found at homepages.caverock.net.nz/˜byfern/list.htm(which can be accessed using the http:// prefix).

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Class Psilotopsida. Insome embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Class Psilotopsida,Order Psilotales. In some embodiments the nucleic acid molecule encodinga PtIP-65 polypeptide is derived from a fern species in the ClassPsilotopsida, Order Ophioglossales. In some embodiments the nucleic acidmolecule encoding a PtIP-65 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales, Family Psilotaceae. Insome embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Class Psilotopsida,Order Ophioglossales Family Ophioglossaceae. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from afern species in the Genus Ophioglossum L., Botrychium, Botrypus,Helminthostachys, Ophioderma, Cheiroglossa, Sceptridium or Mankyua. Insome embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Ophioglossum L. Genusis selected from but not limited to Ophioglossum califomicum,Ophioglossum coriaceum, Ophioglossum costatum, Ophioglossumcrotalophoroides, Ophioglossum engelmannii, Ophioglossum falcaturn,Ophioglossum gomezianurn, Ophioglossum gramineum, Ophioglossumkawamurae, Ophioglossum lusitanicum, Ophioglossum namegatae,Ophioglossum nudicaule, Ophioglossum palmatum, Ophioglossum parvum,Ophioglossum pedunculosum, Ophioglossum pendulum, Ophioglossumpetiolatum, Ophioglossum pusillum, Ophioglossum reticulatum,Ophioglossum richardsiae, Ophioglossum thermale, and Ophioglossumvulgatum.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the ClassPolypodiopsida/Pteridopsida. In some embodiments the nucleic acidmolecule encoding a PtIP-65 polypeptide is derived from a fern speciesin the Order Osmundales (royal ferns); Family Osmundaceae. In someembodiments the nucleic acid molecule encoding a PtIP-65 polypeptide isderived from a fern species in the Order Hymenophyllales (filmy fernsand bristle ferns); Family Hymenophyllaceae. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from afern species in the Order Gleicheniales; Family Gleicheniaceae, FamilyDipteridaceael or Family Matoniaceae. In some embodiments the nucleicacid molecule encoding a PtIP-65 polypeptide is derived from a fernspecies in the Order Schizaeales; Family Lygodiaceae, Family Anemiaceaeor Family Schizaeaceae. In some embodiments the nucleic acid moleculeencoding a PtIP-65 polypeptide is derived from a fern species in theOrder Salviniales; Family Marsileaceae or Family Salviniaceae. In someembodiments the nucleic acid molecule encoding a PtIP-65 polypeptide isderived from a fern species in the Order Cyatheales; FamilyThyrsopteridaceae, Family Loxsomataceae, Family Culcitaceae, FamilyPlagiogyriaceae, Family Cibotiaceae, Family Cyatheaceae, FamilyDicksoniaceae or Family Metaxyaceae.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales;Family Lindsaeaceae, Family Saccolomataceae, Family Cystodiaceae, FamilyDennstaedtiaceae, Family Pteridaceae, Family Aspleniaceae, FamilyThelypteridaceae, Family Woodsiaceae, Family Onocleaceae, FamilyBlechnaceae, Family Dryopteridaceae, Family Lomariopsidaceae, FamilyTectariaceae, Family Oleandraceae, Family Davalliaceae or FamilyPolypodiaceae.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Pteridaceae, Genus Adiantaceae selected from but not limited toAdiantum aethiopicum, Adiantum aleuticum, Adiantum bonatianum, Adiantumcajennense, Adiantum capillus-junonis, Adiantum capillus-veneris,Adiantum caudatum, Adiantum chienfi, Adiantum chilense, Adiantumcuneatum, Adiantum cunninghamfi, Adiantum davidii, Adiantum diaphanum,Adiantum edentulum, Adiantum edgeworthfi, Adiantum excisum, Adiantumfengianum, Adiantum fimbriatum, Adiantum flabellulatum, Adiantumformosanum, Adiantum formosum, Adiantum fulvum, Adiantum gravesfi,Adiantum hispidulum, Adiantum induratum, Adiantum jordanfi, Adiantumjuxtapositum, Adiantum latifolium, Adiantum leveillei, Adiantumlianxianense, Adiantum malesianum, Adiantum mariesfi, Adiantummonochlamys, Adiantum myriosorum, Adiantum obliquum, Adiantumogasawarense, Adiantum pedatum, Adiantum pentadactylon, Adiantumperuvianum, Adiantum philippense, Adiantum princeps, Adiantum pubescens,Adiantum raddianum, Adiantum reniforme, Adiantum roborowskii, Adiantumserratodentatum, Adiantum sinicum, Adiantum soboliferum, Adiantumsubcordatum, Adiantum tenerum, Adiantum terminatum, Adiantumtetraphyllum, Adiantum venustum, Adiantum viridescens, and Adiantumviridimontanum.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Aspleniaceae, Genus Asplenium L. In some embodiments the nucleicacid molecule encoding a PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL selected from but not limited to Asplenium abbreviatum, Aspleniumabrotanoides, Asplenium abscissum var. subaequilaterale, Aspleniumabscissum, Asplenium achilleifolium, Asplenium acuminatum, Aspleniumadiantifrons, Asplenium adiantoides, Asplenium adiantoides var.squamulosum, Asplenium adiantum-nigrum L., Asplenium adiantum-nigrumvar. adiantum-nigrum, Asplenium adiantum-nigrum var. yuanurn, Aspleniumadnatum, Asplenium aethiopicum, Asplenium affine, Asplenium affine var.affine, Asplenium affine var. gilpinae, Asplenium affine var. mettenii,Asplenium affine var. pecten, Asplenium africanum, Asplenium afzelii,Asplenium aitchisonii, Asplenium alatulum, Asplenium alaturn, Aspleniumalfredii, Asplenium altajense, Asplenium amabile, Aspleniumambohitantelense, Asplenium anceps var. proliferum, Asplenium andapense,Asplenium andersonii, Asplenium angustatum, Asplenium angustum,Asplenium anisophyllum, Asplenium annetii, Asplenium antiquum, Aspleniumantrophyoides, Asplenium apertura, Asplenium apogamum, Aspleniumaquaticum, Asplenium arboreum, Asplenium arcanum, Asplenium arcuaturn,Asplenium argentinum, Asplenium argutum, Asplenium aspidiiforme,Asplenium aspidioides, Asplenium asterolepis, Asplenium auriculariumvar. acutidens, Asplenium auricularium var. subintegerrimum, Aspleniumauriculatum, Asplenium auriculatum var. aequilaterale, Asplenium auritumfo. diversifolium, Asplenium auritum fo. diversifolium, Aspleniumauritum fo. nana, Asplenium auritum, Asplenium auritum var. auriculatum,Asplenium auritum var. auritum, Asplenium auritum var. bipinnatifidum,Asplenium auritum var. bipinnatisectum, Asplenium auritum var.davallioides, Asplenium auritum var. macilentum, Asplenium auritum var.rigidum, Asplenium auritum var. subsimplex, Asplenium austrochinense,Asplenium ayopayense, Asplenium badinii, Asplenium balense, Aspleniumballivianii, Asplenium bangii, Asplenium bangii, Asplenium barbaense,Asplenium barclayanum, Asplenium barkamense, Asplenium barteri,Asplenium basiscopicum, Asplenium bicrenatum, Asplenium bifrons,Asplenium bipartitum, Asplenium blastophorum, Asplenium blepharodes,Asplenium blepharophorum, Asplenium boiteaui, Asplenium bolivianurn,Asplenium boltonii, Asplenium borealichinense, Asplenium bradei,Asplenium bradeorum, Asplenium bradleyi, Asplenium brausei, Aspleniumbreedlovei, Asplenium buettneri, Asplenium buettneri var. hildebrandtii,Asplenium bulbiferum, Asplenium bullatum var. bullatum, Aspleniumbullatum var. shikokianum, Asplenium bullatum, Asplenium cancellatum,Asplenium capillipes, Asplenium cardiophyllum (Hance), Aspleniumcaripense, Asplenium carvalhoanurn, Asplenium castaneoviride, Aspleniumcastaneum, Asplenium caudatum, Asplenium celtidifolium (Kunze),Asplenium ceratolepis, Asplenium changputungense, Asplenium chaseanum,Asplenium cheilosorum, Asplenium chengkouense, Asplenium chihuahuense,Asplenium chimantae, Asplenium chimborazense, Asplenium chingianurn,Asplenium chlorophyllum, Asplenium chondrophyllum, Asplenium cicutarium,Asplenium cicutarium var. paleaceum, Asplenium cirrhatum, Aspleniumcladolepton, Asplenium claussenii, Asplenium coenobiale, Aspleniumcommutatum, Asplenium congestum, Asplenium conquisitum, Aspleniumconsimile, Asplenium contiguum, Asplenium contiguum var. hirtulum,Asplenium corderoi, Asplenium cordovense, Asplenium coriaceum, Aspleniumcoriffolium, Asplenium correardii, Asplenium costale, Asplenium costalevar. robustum, Asplenium cowanii, Asplenium crenulatoserrulatum,Asplenium crenulatum, Asplenium crinicaule, Asplenium crinulosum,Asplenium cristatum, Asplenium cryptolepis Fernald, Aspleniumcultrifolium L., Asplenium cuneatiforme, Asplenium cuneaturn, Aspleniumcurvatura, Asplenium cuspidatum, Asplenium cuspidatum var cuspidatum,Asplenium cuspidatum var. foeniculaceum, Asplenium cuspidatum var.triculum, Asplenium cuspidatum var. tripinnaturn, Asplenium dalhousiae,Asplenium dareoides, Asplenium davallioides, Asplenium davisii,Asplenium debile, Asplenium debile, Asplenium decussatum, Aspleniumdelavayi, Asplenium delicatulum, Asplenium delicatulum var. cocosensis,Asplenium delitescens, Asplenium delitescens×laetum, Asplenium densum,Asplenium dentatum L., Asplenium dentatum L., Asplenium depauperaturn,Asplenium deqenense, Asplenium dianae, Asplenium difforme, Aspleniumdilataturn, Asplenium dimidiaturn, Asplenium dimidiatum var. boliviense,Asplenium diplazisorum, Asplenium dissectum, Asplenium distans,Asplenium divaricatum, Asplenium divergens, Asplenium divisissimum,Asplenium doederleinii, Asplenium donnell-smithii, Asplenium dregeanum,Asplenium dulongjiangense, Asplenium duplicatoserratum, Aspleniumeatonii, Asplenium ebeneum, Asplenium ebenoides, Asplenium ecuadorense,Asplenium eggersii, Asplenium emarginatum, Asplenium enaturn, Aspleniumensiforme fo. bicuspe, Asplenium ensiforme fo. ensiforme, Aspleniumensiforme fo. stenophyllum, Asplenium ensiforme, Asplenium erectum var.erectum, Asplenium erectum var. gracile, Asplenium erectum var.usambarense, Asplenium erectum var. zeyheri, &, Asplenium erosum L.,Asplenium escaleroense, Asplenium esculentum, Asplenium eutecnum,Asplenium excelsum, Asplenium excisum, Asplenium exiguum, Aspleniumextensum, Asplenium falcaturn, Asplenium falcinellum, Asplenium faurei,Asplenium feel, Asplenium fengyangshanense, Asplenium ferulaceum,Asplenium fibrillosum, Asplenium filix-femina, Asplenium finckii,Asplenium finlaysonianurn, Asplenium flabellulatum, Aspleniumflabellulatum var flabellulatum, Asplenium flabellulatum var. partitum,Asplenium flaccidum, Asplenium flavescens, Asplenium flavidum, Aspleniumflexuosum, Asplenium fluminense, Asplenium foeniculaceum, Aspleniumformosanum, Asplenium formosum var. carolinurn, Asplenium formosum var.incultum, Asplenium formosum, Asplenium fournieri, Asplenium fragile,Asplenium fragile var. lomense, Asplenium fragrans, Asplenium fragransvar. foeniculaceum, Asplenium franconis var. gracile, Aspleniumfraxinifolium, Asplenium friesiorum, Asplenium friesiorum var.nesophilum, Asplenium fugax, Asplenium fujianense, Asplenium furcatum,Asplenium furfuraceum, Asplenium fuscipes, Asplenium fuscopubescens,Asplenium galeottii, Asplenium gautieri, Asplenium gemmiferum, Aspleniumgentryi, Asplenium geppii, Asplenium ghiesbreghtii, Asplenium gilliesii,Asplenium gilpinae, Asplenium glanduliserraturn, Asplenium glenniei,Asplenium goldmannii, Asplenium gomezianurn, Asplenium grande, Aspleniumgrandifolium, Asplenium grandifrons, Asplenium gregoriae, Aspleniumgriffithianum, Asplenium gulingense, Asplenium hainanense, Aspleniumhallbergii, Asplenium hallei, Asplenium hallii, Asplenium hangzhouense,Asplenium haplophyllum, Asplenium harpeodes, Asplenium harpeodes var.glaucovirens, Asplenium harpeodes var. incisura, Asplenium harrisiiJenman, Asplenium harrisonii, Asplenium hastatum, Asplenium hebeiense,Asplenium hemionitideum, Asplenium hemitomurn, Asplenium hentyi,Asplenium herpetopteris, Asplenium herpetopteris var herpetopteris,Asplenium herpetopteris var. acutipinnata, Asplenium herpetopteris var.masoulae, Asplenium herpetopteris var. villosum, Asplenium hesperium,Asplenium heterochroum, Asplenium hians, Asplenium hians var.pallescens, Asplenium hoffmannii, Asplenium holophlebium, Aspleniumhondoense, Asplenium horridum, Asplenium hostmannii, Aspleniumhumistratum, Asplenium hypomelas, Asplenium inaequilaterale, Aspleniumincisura, Asplenium incurvaturn, Asplenium indicum, Asplenium indicumvar. indicum, Asplenium indicum var. yoshingagae, Asplenium induratum,Asplenium indusiatum, Asplenium inexpectatum, Asplenium insigne,Asplenium insiticium, Asplenium insolitum, Asplenium integerrimurn,Asplenium interjectum, Asplenium jamesonii, Asplenium jaundeense,Asplenium juglandifolium, Asplenium kangdingense, Asplenium kansuense,Asplenium kassneri, Asplenium kaulfussii, Asplenium kellermanii,Asplenium kentuckiense, Asplenium khullarii, Asplenium kiangsuense,Asplenium kunzeanurn, Asplenium lacerum, Asplenium laciniatum, Aspleniumlaciniatum var. acutipinna, Asplenium laciniatum var. laciniatum,Asplenium laetum fo. minor, Asplenium laetum, Asplenium laetum var.incisoserratum, Asplenium lamprocaulon, Asplenium laserpitiifolium var.morrisonense, Asplenium lastii, Asplenium latedens, Aspleniumlatifolium, Asplenium laui, Asplenium laurentii, Asplenium leandrianurn,Asplenium lechleri, Asplenium leiboense, Asplenium lepidorachis,Asplenium leptochlamys, Asplenium leptophyllum, Asplenium levyi,Asplenium lindbergii, Asplenium lindeni, Asplenium lineatum, Aspleniumlividum, Asplenium lobatum, Asplenium lobulaturn, Asplenium lokohoense,Asplenium longicauda, Asplenium longicaudaturn, Asplenium longifolium,Asplenium longisorum, Asplenium longjinense, Asplenium lorentzii,Asplenium loriceum, Asplenium loxogrammoides, Asplenium lugubre,Asplenium lunulatum, Asplenium lunulatum var. pteropus, Aspleniumlushanense, Asplenium lydgatei, Asplenium macilentum, Asplenium macraei,Asplenium macrodictyon, Asplenium macrophlebium, Asplenium macrophyllum,Asplenium macropterum, Asplenium macrosorum, Asplenium macrotis,Asplenium macrurum, Asplenium mainlingense, Asplenium mangindranense,Asplenium mannii, Asplenium marginatum L., Asplenium marojejyense,Asplenium martianum, Asplenium matsumurae, Asplenium mauritiensisLorence, Asplenium maximum, Asplenium, ii, Asplenium megalura, Aspleniummegaphyllum, Asplenium meiotomurn, Asplenium melanopus, Aspleniummembranifolium, Asplenium meniscioides, Asplenium mesosorum, Aspleniummexicanurn, Asplenium micropaleatum, Asplenium microtum, Aspleniummildbraedii, Asplenium mildei, Asplenium minimum, Asplenium minutum,Asplenium miradorense, Asplenium miyunense, Asplenium moccenianurn,Asplenium mocquetysii, Asplenium modestum, Asplenium monanthemum var.menziesii, Asplenium monanthes L., Asplenium monanthes var monanthes,Asplenium monanthes var. castaneum, Asplenium monanthes var. wagneri,Asplenium monanthes var. yungense, Asplenium monodon, Aspleniummontanum, Asplenium mosetenense, Asplenium moupinense, Aspleniummucronatum, Asplenium munchii, Asplenium muticum, Asplenium myapteron,Asplenium myriophyllu, Asplenium nakanoanum, Asplenium nanchuanense,Asplenium nemorale, Asplenium neolaserpitiifolium, Aspleniumneomutijugum, Asplenium neovarians, Asplenium nesii, Aspleniumnesioticum, Asplenium nidus L., Asplenium nigricans, Aspleniumniponicum, Asplenium normale, Asplenium normale var. angustum, Aspleniumobesum, Asplenium oblongatum, Asplenium oblongifolium, Aspleniumobovatum, Asplenium obscurum, Asplenium obscurum var. angustum,Asplenium obtusatum var. obtusatum, Asplenium obtusatum var. sphenoides,Asplenium obtusifolium L., Asplenium obtusissimum, Asplenium obversum,Asplenium ochraceum, Asplenium oellgaardii, Asplenium ofeliae, Aspleniumoldhami, Asplenium oligosorum, Asplenium olivaceum, Asplenium onopterisL., Asplenium onustum, Asplenium ortegae, Asplenium otites, Aspleniumpalaciosii, Asplenium palmeri, Asplenium partitum, Asplenium parvisorum,Asplenium parviusculum, Asplenium parvulum, Asplenium patens, Aspleniumpaucifolium, Asplenium paucijugum, Asplenium paucivenosum, Aspleniumpearcei, Asplenium pekinense, Asplenium pellucidum, Asplenium pendulum,Asplenium petiolulatum, Asplenium phyllitidis, Aspleniumpimpinellifolium, Asplenium pinnatifidum, Asplenium pinnatum, Aspleniumplatyneuron, Asplenium platyneuron var. bacculum-rubrum, Aspleniumplatyneuron var. incisum, Asplenium platyphyllum, Asplenium plumbeum,Asplenium poloense, Asplenium polymeris, Asplenium polymorphum,Asplenium polyodon, Asplenium polyodon var. knudsenii, Aspleniumpolyodon var. nitidulum, Asplenium polyodon var. sectum, Aspleniumpolyodon var. subcaudatum, Asplenium polyphyllum, Asplenium poolii,Asplenium poolii fo. simplex, Asplenium poolii var. linearipinnatum,Asplenium potosinum, Asplenium potosinum var. incisum, Aspleniumpraegracile, Asplenium praemorsum, Asplenium preussii, Aspleniumpringleanum, Asplenium pringlei, Asplenium prionitis, Aspleniumprocerum, Asplenium progrediens, Asplenium projectum, Aspleniumprolongatum, Asplenium propinquum, Asplenium protensum, Aspleniumpseudoangustum, Asplenium pseudoerectum, Asplenium pseudofontanum,Asplenium pseudolaserpitiifolium, Asplenium pseudonormale, Aspleniumpseudo pellucidum, Asplenium pseudopraemorsum, Asplenium pseudovarians,Asplenium pseudowilfordii, Asplenium pseudowrightii, Aspleniumpsilacrum, Asplenium pteropus, Asplenium pubirhizoma, Aspleniumpulchellum, Asplenium pulchellum var. subhorizontale, Aspleniumpulcherrimum, Asplenium pulicosum, Asplenium pulicosum var. maius,Asplenium pululahuae, Asplenium pumilum, Asplenium pumilum var.hymenophylloides, Asplenium pumilum var. laciniatum, Aspleniumpurdieanum, Asplenium purpurascens, Asplenium pyramidatum, Aspleniumqiujiangense, Asplenium quercicola, Asplenium quitense, Aspleniumraddianum, Asplenium radiatum, Asplenium radicans L., Aspleniumradicans, Asplenium radicans var. costaricense, Asplenium radicans var.partitum, Asplenium radicans var. radicans, Asplenium radicans var.uniseriale, Asplenium recumbens, Asplenium reflexum, Asplenium regularevar. latior, Asplenium repandulum, Asplenium repens, Asplenium repente,Asplenium resiliens, Asplenium retusulum, Asplenium rhipidoneuron,Asplenium rhizophorum L., Asplenium rhizophyllum, Asplenium rhizophyllumL., Asplenium rhizophyllum var. proliferum, Asplenium rhomboideum,Asplenium rigidum, Asplenium riparium, Asplenium rivale, Aspleniumrockii, Asplenium roemerianum, Asplenium roemerianum var. mindensis,Asplenium rosenstockianum, Asplenium rubinum, Asplenium ruizianum,Asplenium rusbyanum, Asplenium ruta-muraria L., Asplenium ruta-murariavar. cryptolepis, Asplenium rutaceum, Asplenium rutaceum var.disculiferum, Asplenium rutaefolium, Asplenium rutifolium, Aspleniumsalicifolium L., Asplenium salicifolium var. aequilaterale, Aspleniumsalicifolium var. salicifolium, Asplenium sampsoni, Asplenium sanchezii,Asplenium sanderi, Asplenium sandersonii, Asplenium sanguinolentum,Asplenium sarelii, Asplenium sarelii var. magnum, Asplenium sarelii var.sarelii, Asplenium saxicola, Asplenium scalifolium, Aspleniumscandicinum, Asplenium schizophyllum, Asplenium schkuhrii, Aspleniumsciadophilum, Asplenium scolopendrium L., Asplenium scortechinii,Asplenium seileri, Asplenium semipinnatum, Asplenium septentrionale,Asplenium serra, Asplenium serra var. imrayanum, Aspleniumserratissimum, Asplenium serratum L., Asplenium serratum var. caudatum,Asplenium serricula, Asplenium sessilifolium, Asplenium sessilifoliumvar. guatemalense, Asplenium sessilifolium var. minus, Aspleniumsessilifolium var. occidentale, Asplenium sessilipinnum, Aspleniumsetosum, Asplenium shepherdii, Asplenium shepherdii var. bipinnatum,Asplenium shepherdii var. flagelliferum, Asplenium shikokianum,Asplenium simii, Asplenium simonsianum, Asplenium sintenisii, Aspleniumskinneri, Asplenium skinneri, Asplenium sodiroi, Aspleniumsoleirolioides, Asplenium solidum var. stenophyllum, Asplenium solmsii,Asplenium sp.-N.-Halle-2234, Asplenium spathulinum, Aspleniumspectabile, Asplenium speluncae, Asplenium sphaerosporum, Aspleniumsphenotomum, Asplenium spinescens, Asplenium splendens, Aspleniumsprucei, Asplenium squamosum L., Asplenium standleyi, Aspleniumstellatum, Asplenium stenocarpum, Asplenium stoloniferum, Aspleniumstolonipes, Asplenium striatum L., Asplenium stuebelianum, Aspleniumstuhlmannii, Asplenium suave, Asplenium subalatum, Aspleniumsubcrenatum, Asplenium subdigitatum, Asplenium subdimidiatum, Aspleniumsubintegrum, Asplenium sublaserpitiifolium, Asplenium sublongum,Asplenium subnudum, Asplenium suborbiculare, Asplenium subtenuifolium,Asplenium subtile, Asplenium subtoramanum, Asplenium subtrapezoideum,Asplenium subvarians, Asplenium sulcatum, Asplenium sylvaticum,Asplenium szechuanense, Asplenium taiwanense, Asplenium tenerrimum,Asplenium tenerum, Asplenium tenuicaule, Asplenium tenuifolium,Asplenium tenuifolium var. minor, Asplenium tenuifolium var.tenuifolium, Asplenium tenuissimum, Asplenium ternatum, Aspleniumtheciferum, Asplenium theciferum var. concinnum, Asplenium thunbergii,Asplenium tianmushanense, Asplenium tianshanense, Asplenium tibeticum,Asplenium tocoraniense, Asplenium toramanum, Asplenium trapezoideum,Asplenium tricholepis, Asplenium trichomanes L., Asplenium trichomanessubsp. inexpectans, Asplenium trichomanes subsp. quadrivalens, Aspleniumtrichomanes subsp. trichomanes, Asplenium trichomanes var. harovii,Asplenium trichomanes var. herbaceum, Asplenium trichomanes var. repens,Asplenium trichomanes var. viridissimum, Asplenium trichomanes-dentatumL., Asplenium trigonopterum, Asplenium trilobatum, Asplenium trilobum,Asplenium triphyllum, Asplenium triphyllum var. cornpactum, Aspleniumtriphyllum var. gracillimum, Asplenium triphyllum var. herbaceum,Asplenium tripteropus, Asplenium triquetrum, Asplenium truncorum,Asplenium tsaratananense, Asplenium tucumanense, Asplenium tuerckheimii,Asplenium tunquiniense, Asplenium ulbrichtii, Asplenium ultimurn,Asplenium unilaterale, Asplenium unilaterale var. decurrens, Aspleniumunilaterale var. udum, Asplenium unilaterale var. unilaterale, Aspleniumuniseriale, Asplenium uropteron, Asplenium vagans, Aspleniumvareschianum, Asplenium variabile var. paucijugum, Asplenium variabilevar. variabile, Asplenium varians subsp. fimbriatum, Asplenium varians,Asplenium vastum, Asplenium venturae, Asplenium venulosum, Aspleniumverapax, Asplenium vesiculosum, Asplenium vespertinurn, Aspleniumvillosum, Asplenium virens, Asplenium viride, Asplenium viridifrons,Asplenium virillae, Asplenium viviparioides, Asplenium viviparum,Asplenium viviparum var viviparum, Asplenium viviparum var. lineatu,Asplenium volubile, Asplenium vulcanicum, Asplenium wacketii, Aspleniumwagneri, Asplenium wallichianum, Asplenium warneckei, Aspleniumwilfordii, Asplenium williamsii, Asplenium wrightii, Aspleniumwrightioides, Asplenium wuliangshanense, Asplenium xianqianense,Asplenium xinjiangense, Asplenium xinyiense, Asplenium yelagagense,Asplenium yoshinagae, Asplenium yunnanense, Asplenium zamiifolium,Asplenium zanzibaricum, Asplenium biscayneanum, Asplenium curtissii,Asplenium ebenoides, Asplenium herb-wagneri, Asplenium heteroresiliens,Asplenium kenzoi, Asplenium plenum, Asplenium wangii, andAsplenium×clermontiae, Asplenium×gravesii.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Blechnaceae, Genus Blechnum L. In some embodiments the nucleicacid molecule encoding a PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.selected from but not limited to Blechnum amabile, Blechnumappendiculatum, Blechnum articulatum, Blechnum australe, Blechnumaustrobrasilianum, Blechnum binervatum, Blechnum blechnoides, Blechnumbrasiliense, Blechnum capense, Blechnum cartilagineum, Blechnumcastaneum, Blechnum chambersii, Blechnum chilense, Blechnum colensoi,Blechnum contiguum, Blechnum cordatum, Blechnum coriaceum, Blechnumdiscolor, Blechnum doodioides, Blechnum durum, Blechnum eburneum,Blechnum ensiforme, Blechnum filiforme, Blechnum fluviatile, Blechnumfragile, Blechnum fraseri, Blechnum fullagari, Blechnum gibbum, Blechnumglandulosum, Blechnum gracile, Blechnum hancockii, Blechnum hastatum,Blechnum howeanum, Blechnum indicum, Blechnum kunthianum, Blechnumlaevigatum, Blechnum loxense, Blechnum magellanicum, Blechnummembranaceum, Blechnum microbasis, Blechnum microphyllum, Blechnummilnei, Blechnum minus, Blechnum mochaenum, Blechnum montanum, Blechnummoorei, Blechnum moritzianum, Blechnum nigrum, Blechnum niponicum,Blechnum norfolkianum, Blechnum novae-zelandiae, Blechnum nudurn,Blechnum obtusatum, Blechnum occidentale, Blechnum oceanicum, Blechnumorientale, Blechnum patersonii, Blechnum penna-marina, Blechnumpolypodioides, Blechnum procerum, Blechnum punctulatum, Blechnumsampaioanum, Blechnum schiedeanum, Blechnum schomburgkii, Blechnumserrulatum, Blechnum simillimum, Blechnum spicant, Blechnumstipitellatum, Blechnum tabulare, Blechnum triangularifolium, Blechnumvieillardii, Blechnum vulcanicum, Blechnum wattsii, Blechnum whelanii,and Blechnum wurunuran.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Dryopteridaceae Genus Acrophorus, Genus Acrorumohra, GenusAnapausia, Genus Arachniodes, Genus Bolbitis, Genus Ctenitis, GenusCyclodium, Genus Cyrtogonellum, Genus Cyrtomidictyum, Genus Cyrtomium,Genus Diacalpe, Genus Didymochlaena, Genus Dryopsis, Genus Dryopteris,Genus Elaphoglossum, Genus Hypodematium, Genus Lastreopsis, GenusLeptorumohra, Genus Leucostegia, Genus Lithostegia, Genus Lomagramma,Genus Maxonia, Genus Megalastrum, Genus Olfersia, Genus Peranema, GenusPhanerophlebia, Genus Phanerophlebiopsis, Genus Polybotrya, GenusPolystichopsis, Genus Polystichum, Genus Rumohra, Genus Sorolepidium,Genus Stigmatopteris or Genus Teratophyllum. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from afern species in the Order Polypodiales, Family Dryopteridaceae, GenusBolbitis, selected from but not limited to Bolbitis acrostichoides,Bolbitis aliena, Bolbitis angustipinna, Bolbitis appendiculata, Bolbitisauriculata, Bolbitis bernoullii, Bolbitis bipinnatifida, Bolbitiscadieri, Bolbitis christensenii, Bolbitis confertifolia, Bolbitiscostata, Bolbitis crispatula, Bolbitis fluviatilis, Bolbitisgaboonensis, Bolbitis gemmifera, Bolbitis hainanensis, Bolbitis hastata,Bolbitis hekouensis, Bolbitis hemiotis, Bolbitis heteroclita, Bolbitisheudelotii, Bolbitis humblotii, Bolbitis interlineata, Bolbitislatipinna, Bolbitis laxireticulata, Bolbitis lindigii, Bolbitislonchophora, Bolbitis longiflagellata, Bolbitis major, Bolbitis media,Bolbitis nicotianifolia, Bolbitis nodiflora, Bolbitis novoguineensis,Bolbitis oligarchica, Bolbitis palustris, Bolbitis pandurifolia,Bolbitis pergamentacea, Bolbitis portoricensis, Bolbitis presliana,Bolbitis quoyana, Bolbitis rawsonii, Bolbitis repanda, Bolbitisrhizophylla, Bolbitis riparia, Bolbitis rivularis, Bolbitis sagenioides,Bolbitis salicina, Bolbitis scalpturata, Bolbitis scandens, Bolbitissemicordata, Bolbitis semipinnatifida, Bolbitis serrata, Bolbitisserratifolia, Bolbitis simplex, Bolbitis sinensis, Bolbitissingaporensis, Bolbitis sinuata, Bolbitis subcordata, Bolbitissubcrenata, Bolbitis taylorii, Bolbitis tibetica, Bolbitis tonkinensis,Bolbitis umbrosa, Bolbitis vanuaensis, and Bolbitis virens.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Lomariopsidaceae, Genus Nephrolepis. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from afern species in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis is selected from but not limited to Nephrolepis abrupta,Nephrolepis acuminata, Nephrolepis acutifolia, Nephrolepis arida,Nephrolepis arthropteroides, Nephrolepis biserrata var. auriculata,Nephrolepis brownii, Nephrolepis celebica, Nephrolepis clementis,Nephrolepis cordifolia, Nephrolepis davalliae, Nephrolepis davallioides,Nephrolepis dayakorum, Nephrolepis delicatula, Nephrolepisdicksonioides, Nephrolepis duffii, Nephrolepis exaltata ssp. exaltatassp. Hawaiiensis, Nephrolepis falcate, Nephrolepis falciformis,Nephrolepis glabra, Nephrolepis hirsutula, Nephrolepis humatoides,Nephrolepis iridescens, Nephrolepis kurotawae, Nephrolepis laurifolia,Nephrolepis lauterbachii, Nephrolepis lindsayae, Nephrolepis multifida,Nephrolepis multiflora, Nephrolepis niphoboloides, Nephrolepisobliterate, Nephrolepis paludosa, Nephrolepis pectinate, Nephrolepispendula, Nephrolepis persicifolia, Nephrolepis pickelii, Nephrolepispilosula, Nephrolepis pubescens, Nephrolepis pumicicola, Nephrolepisradicans, Nephrolepis rivularis, Nephrolepis rosenstockii, Nephrolepissaligna, Nephrolepis schlechteri, Nephrolepis serrate, Nephrolepisthomsoni, Nephrolepis undulate var. aureoglandulosa, Nephrolepis×averyi,Nephrolepis×copelandii, and Nephrolepis×medlerae.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Polypodiaceae Genus Campyloneurum, Genus Drynaria, GenusLepisorus, Genus Microgramma, Genus Microsorum, Genus Neurodium, GenusNiphidium, Genus Pecluma M.G., Genus Phlebodium, Genus Phymatosorus,Genus Platycerium, Genus Pleopeltis, Genus Polypodium L.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a fern species in the Order Polypodiales,Family Polypodiaceae, Genus Polypodium L. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from afern species in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L. selected from but not limited to Polypodium absidatum,Polypodium acutifolium, Polypodium adiantiforme, Polypodium aequale,Polypodium affine, Polypodium albidopaleatum, Polypodium alcicorne,Polypodium alfarii, Polypodium alfredii, Polypodium alfredii var.curtii, Polypodium allosuroides, Polypodium alsophilicola, Polypodiumamamianum, Polypodium amoenum, Polypodium amorphum, Polypodiumanetioides, Polypodium anfractuosum, Polypodium anguinum, Polypodiumangustifolium f. remotifolia, Polypodium angustifolium var. amphostenon,Polypodium angustifolium var. heterolepis, Polypodium angustifolium var.monstrosa, Polypodium angustipaleatum, Polypodium angustissimum,Polypodium anisomeron var. pectinatum, Polypodium antioquianum,Polypodium aoristisorum, Polypodium apagolepis, Polypodium apicidens,Polypodium apiculatum, Polypodium apoense, Polypodium appalachianum,Polypodium appressum, Polypodium arenarium, Polypodium argentinum,Polypodium argutum, Polypodium armaturn, Polypodium aromaticum,Polypodium aspersum, Polypodium assurgens, Polypodium atrum, Polypodiumauriculatum, Polypodium balaonense, Polypodium balliviani, Polypodiumbamleri, Polypodium bangii, Polypodium bartlettii, Polypodium basale,Polypodium bemoullii, Polypodium biauritum, Polypodium bifrons,Polypodium blepharodes, Polypodium bolivari, Polypodium bolivianum,Polypodium bolobense, Polypodium bombycinum, Polypodium bombycinum var.insularum, Polypodium bradeorum, Polypodium bryophilum, Polypodiumbryopodum, Polypodium buchtienii, Polypodium buesii, Polypodiumbulbotrichum, Polypodium caceresii, Polypodium californicum f.brauscombii, Polypodium californicum f. parsonsiae, Polypodiumcalifornicum, Polypodium calophlebium, Polypodium calvum, Polypodiumcamptophyllarium var. abbreviatum, Polypodium capitellatum, Polypodiumcarpinterae, Polypodium chachapoyense, Polypodium chartaceum, Polypodiumchimantense, Polypodium chiricanum, Polypodium choquetangense,Polypodium christensenii, Polypodium christii, Polypodium chrysotrichum,Polypodium ciliolepis, Polypodium cinerascens, Polypodium collinsii,Polypodium colysoides, Polypodium confluens, Polypodium conforme,Polypodium confusum, Polypodium congregatifolium, Polypodium connellii,Polypodium consimile var. bourgaeanum, Polypodium consimile var. minor,Polypodium conterminans, Polypodium contiguum, Polypodium cookii,Polypodium coriaceum, Polypodium coronans, Polypodium costaricense,Polypodium costatum, Polypodium crassifolium f. angustissimum,Polypodium crassifolium var. longipes, Polypodium crassulum, Polypodiumcraterisorum, Polypodium cryptum, Polypodium crystalloneuron, Polypodiumcucullaturn var. planum, Polypodium cuencanum, Polypodium cumingianum,Polypodium cupreolepis, Polypodium curranii, Polypodium curvans,Polypodium cyathicola, Polypodium cyathisorum, Polypodium cyclocolpon,Polypodium daguense, Polypodium damunense, Polypodium dareiformioides,Polypodium dasypleura, Polypodium decipiens, Polypodium decorum,Polypodium delicatulum, Polypodium deltoideum, Polypodium demeraranum,Polypodium denticulatum, Polypodium diaphanum, Polypodium dilatatum,Polypodium dispersum, Polypodium dissectum, Polypodium dissimulans,Polypodium dolichosorum, Polypodium dolorense, Polypodiumdonnell-smithii, Polypodium drymoglossoides, Polypodium ebeninum,Polypodium eggersii, Polypodium elmeri, Polypodium elongatum, Polypodiumenterosoroides, Polypodium erubescens, Polypodium erythrolepis,Polypodium erythrotrichum, Polypodium eurybasis, Polypodium eurybasisvar. villosum, Polypodium exornans, Polypodium falcoideum, Polypodiumfallacissimum, Polypodium farinosum, Polypodium faucium, Polypodium feetPolypodium ferrugineum, Polypodium feuillei, Polypodium firmulum,Polypodium firmum, Polypodium flaccidum, Polypodium flagellare,Polypodium flexuosum, Polypodium flexuosum var. ekmanii, Polypodiumforbesii, Polypodium formosanum, Polypodium fraxinifolium subsp.articulatum, Polypodium fraxinifolium subsp. luridum, Polypodiumfructuosum, Polypodium fucoides, Polypodium fulvescens, Polypodiumgaleottii, Polypodium glaucum, Polypodium glycyrrhiza, Polypodiumgracillimum, Polypodium gramineum, Polypodium grandifolium, Polypodiumgratum, Polypodium graveolens, Polypodium griseo-nigrum, Polypodiumgriseum, Polypodium guttatum, Polypodium haalilioanum, Polypodiumhammatisorum, Polypodium hancockii, Polypodium haplophlebicum,Polypodium harrisii, Polypodium hastatum var. simplex, Polypodiumhawaiiense, Polypodium heanophyllum, Polypodium helleri, Polypodiumhemionitidium, Polypodium henryi, Polypodium herzogii, Polypodiumhesperium, Polypodium hessii, Polypodium hombersleyi, Polypodiumhostmannii, Polypodium humile, Polypodium hyalinum, Polypodium iboense,Polypodium induens var. subdentatum, Polypodium insidiosum, Polypodiuminsigne, Polypodium intermedium subsp. masafueranum var. obtuseserratum,Polypodium intramarginale, Polypodium involutum, Polypodium itatiayense,Polypodium javanicum, Polypodium juglandifolium, Polypodium kaniense,Polypodium knowltoniorum, Polypodium kyimbilense, Polypodiuml'herminieri var. costaricense, Polypodium lachniferum f. incurvata,Polypodium lachniferum var. glabrescens, Polypodium lachnopus,Polypodium lanceolatum var. complanatum, Polypodium lanceolatum var.trichophorum, Polypodium latevagans, Polypodium laxifrons, Polypodiumlaxifrons var. lividum, Polypodium lehmannianum, Polypodium leiorhizum,Polypodium leptopodon, Polypodium leuconeuron var. angustifolia,Polypodium leuconeuron var. latifolium, Polypodium leucosticta,Polypodium limulum, Polypodium lindigii, Polypodium lineatum, Polypodiumlomarioides, Polypodium longifrons, Polypodium loretense, Polypodiumloriceum var. umbraticum, Polypodium loriforme, Polypodium loxogramme f.gigas, Polypodium ludens, Polypodium luzonicum, Polypodium lycopodioidesf. obtusum, Polypodium lycopodioides L., Polypodium macrolepis,Polypodium macrophyllum, Polypodium macrosorum, Polypodiummacrosphaerum, Polypodium maculosum, Polypodium madrense, Polypodiummanmeiense, Polypodium margaritiferum, Polypodium maritimum, Polypodiummartensii, Polypodium mayoris, Polypodium megalolepis, Polypodiummelanotrichum, Polypodium menisciifolium var. pubescens, Polypodiummeniscioides, Polypodium merrillii, Polypodium mettenii, Polypodiummexiae, Polypodium microsorum, Polypodium militare, Polypodium minimum,Polypodium minusculum, Polypodium mixtum, Polypodium mollendense,Polypodium mollissimum, Polypodium moniliforme var. minus, Polypodiummonoides, Polypodium monticola, Polypodium montigenum, Polypodiummoritzianum, Polypodium moultonii, Polypodium multicaudatum, Polypodiummultilineatum, Polypodium multisorum, Polypodium munchii, Polypodiummuscoides, Polypodium myriolepis, Polypodium myriophyllum, Polypodiummyriotrichum, Polypodium nematorhizon, Polypodium nemorale, Polypodiumnesioticum, Polypodium nigrescentium, Polypodium nigripes, Polypodiumnigrocinctum, Polypodium nimbatum, Polypodium nitidissimum, Polypodiumnitidissimum var. latior, Polypodium nubrigenum, Polypodium oligolepis,Polypodium oligosorum, Polypodium oligosorum, Polypodium olivaceum,Polypodium olivaceum var. elatum, Polypodium oodes, Polypodiumoosphaerum, Polypodium oreophilum, Polypodium ornatissimum, Polypodiumornatum, Polypodium ovatum, Polypodium oxylobum, Polypodium oxypholis,Polypodium pakkaense, Polypodium pallidum, Polypodium palmatopedatum,Polypodium palmeri, Polypodium panamense, Polypodium parvum, Polypodiumpatagonicum, Polypodium paucisorum, Polypodium pavonianum, Polypodiumpectinatum var. caliense, Polypodium pectinatum var. hispidum,Polypodium pellucidum, Polypodium pendulum var. boliviense, Polypodiumpercrassum, Polypodium perpusillum, Polypodium peruvianum var.subgibbosum, Polypodium phyllitidis var. elongatum, Polypodiumpichinchense, Polypodium pilosissimum, Polypodium pilosissimum var.glabriusculum, Polypodium pilossimum var. tunguraquensis, Polypodiumpityrolepis, Polypodium platyphyllum, Polypodium playfairii, Polypodiumplebeium var. cooperi, Polypodium plectolepidioides, Polypodiumpleolepis, Polypodium plesiosorum var.i, Polypodium podobasis,Polypodium podocarpum, Polypodium poloense, Polypodium polydatylon,Polypodium polypodioides var. aciculare, Polypodium polypodioides var.michauxianum, Polypodium praetermissum, Polypodium preslianum var.immersum, Polypodium procerum, Polypodium procerum, Polypodiumproductum, Polypodium productum, Polypodium prolongilobum, Polypodiumpropinguum, Polypodium proteus, Polypodium pruinatum, Polypodiumpseudocapillare, Polypodium pseudofratemum, Polypodium pseudonutans,Polypodium pseudoserratum, Polypodium pulcherrimurn, Polypodiumpulogense, Polypodium pungens, Polypodium purpusii, Polypodium radicale,Polypodium randallii, Polypodium ratiborii, Polypodium reclinaturn,Polypodium recreense, Polypodium repens var. abruptum, Polypodiumrevolvens, Polypodium rhachipterygium, Polypodium rhomboideum,Polypodium rigens, Polypodium robustum, Polypodium roraimense,Polypodium roraimense, Polypodium rosei, Polypodium rosenstockii,Polypodium rubidum, Polypodium rudimentum, Polypodium rusbyi, Polypodiumsablanianum, Polypodium sarmentosum, Polypodium saxicola, Polypodiumschenckii, Polypodium schlechteri, Polypodium scolopendria, Polypodiumscolopendria, Polypodium scolopendrium, Polypodium scouleri, Polypodiumscutulatum, Polypodium segregatum, Polypodium semihirsutum, Polypodiumsemihirsutum var. fuscosetosum, Polypodium senile var. minor, Polypodiumsericeolanatum, Polypodium serraeforme, Polypodium serricula, Polypodiumsesquipedala, Polypodium sessilifolium, Polypodium setosum var. calvum,Polypodium setulosum, Polypodium shaferi, Polypodium sibomense,Polypodium siccum, Polypodium simacense, Polypodium simulans, Polypodiumsingeri, Polypodium sinicum, Polypodium sintenisii, Polypodium skutchii,Polypodium sloanei, Polypodium sodiroi, Polypodium sordidulum,Polypodium sordidum, Polypodium sphaeropteroides, Polypodium sphenodes,Polypodium sprucei, Polypodium sprucei var. furcativenosa, Polypodiumsteirolepis, Polypodium stenobasis, Polypodium stenolepis, Polypodiumstenopterum, Polypodium subcapillare, Polypodium subflabelliforme,Polypodium subhemionitidium, Polypodium subinaequale, Polypodiumsubintegrum, Polypodium subspathulatum, Polypodium subtile, Polypodiumsubvestitum, Polypodium subviride, Polypodium superficiale var.attenuatum, Polypodium superficiale var. chinensis, Polypodiumsursumcurrens, Polypodium tablazianurn, Polypodium taenifolium,Polypodium tamandarei, Polypodium tatei, Polypodium tenuiculum var.acrosora, Polypodium tenuiculum var. brasiliense, Polypodium tenuilore,Polypodium tenuinerve, Polypodium tepuiense, Polypodium teresae,Polypodium tetragonum var. incompletum, Polypodium thysanolepis var.bipinnatifidum, Polypodium thyssanolepis, var. thyssanolepis, Polypodiumthyssanolepsi, Polypodium tobagense, Polypodium trichophyllum,Polypodium tridactylum, Polypodium tridentatum, Polypodium trifurcaturnvar. brevipes, Polypodium triglossum, Polypodium truncatulum, Polypodiumtruncicola var. major, Polypodium truncicola var. minor, Polypodiumtuberosum, Polypodium tunguraguae, Polypodium turquinurn, Polypodiumturrialbae, Polypodium ursipes, Polypodium vagans, Polypodiumvaldealaturn, Polypodium versteegii, Polypodium villagranii, Polypodiumvirginianum f. cambroideum, Polypodium virginianurn f. peraferens,Polypodium vittarioides, Polypodium vulgare, Polypodium vulgare L.,Polypodium vulgare subsp. oreophilum, Polypodium vulgare var.acuminatum, Polypodium vulpinum, Polypodium williamsii, Polypodiumwobbense, Polypodium×fallacissimum-guttatum, Polypodium xantholepis,Polypodium xiphopteris, Polypodium yarumalense, Polypodium yungense, andPolypodium zosteriforme.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium. In some embodiments the PtIP-65 polypeptide is derived froma fern species in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium selected from but not limited to Platycerium alcicorne,Platycerium andinum, Platycerium angolense, Platycerium bifurcatum,Platycerium coronarium, Platycerium elephantotis, Platycerium ellisii,Platycerium grande, Platycerium hillii, Platycerium holtturnii,Platycerium madagascariense, Platycerium quadridichotomum, Platyceriumridleyi, Platycerium sp. ES-2011, Platycerium stemaria, Platyceriumsuperbum, Platycerium veitchii, Platycerium wallichii, Platyceriumwandae, Platycerium wilhelminae-reginae, and Platycerium willinckii.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the Division Lycophyta.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the Class Isoetopsida or ClassLycopodiopsida. In some embodiments the nucleic acid molecule encoding aPtIP-65 polypeptide is derived from a species in the Class IsoetopsidaOrder Selaginales. In some embodiments the nucleic acid moleculeencoding a PtIP-65 polypeptide is derived from a fern species in theClass Isoetopsida, Order Selaginales, Family Selaginellaceae. In someembodiments the nucleic acid molecule encoding a PtIP-65 polypeptide isderived from a species in the Genus Selaginella. In some embodiments thenucleic acid molecule encoding a PtIP-65 polypeptide is derived from aSelaginella species selected from but not limited to Selaginellaacanthonota, Selaginella apoda, Selaginella arbuscula, Selaginellaarenicola, Selaginella arizonica, Selaginella armata, Selaginellaasprella, Selaginella biformis, Selaginella bigelovii, Selaginellabraunii, Selaginella cinerascens, Selaginella cordifolia, Selaginelladeflexa, Selaginella delicatula, Selaginella densa, Selaginelladouglasii, Selaginella eatonii, Selaginella eclipes, Selaginellaeremophila, Selaginella erythropus, Selaginella flabellata, Selaginellahansenii, Selaginella heterodonta, Selaginella kraussiana, Selaginellakrugii, Selaginella laxifolia, Selaginella lepidophylla, Selaginellaleucobryoides, Selaginella ludoviciana, Selaginella mutica, Selaginellaoregana, Selaginella ovifolia, Selaginella pallescens, Selaginellaperuviana, Selaginella pilifera, Selaginella plana, Selaginella plumosa,Selaginella pulcherrima, Selaginella rupestris, Selaginella rupincola,Selaginella scopulorum, Selaginella selaginoides, Selaginella sibirica,Selaginella standleyi, Selaginella stellata, Selaginella subcaulescens,Selaginella substipitata, Selaginella tenella, Selaginella tortipila,Selaginella uliginosa, Selaginella umbrosa, Selaginella uncinata,Selaginella underwoodii, Selaginella utahensis, Selaginella victoriae,Selaginella viridissima, Selaginella wallacei, Selaginella watsonii,Selaginella weatherbiana, Selaginella willdenowii, Selaginella wrightiiand Selaginella×neomexicana.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the Class Lycopodiopsida, OrderLycopodiales. In some embodiments the nucleic acid molecule encoding aPtIP-65 polypeptide is derived from a fern species in the ClassLycopodiopsida, Order Lycopodiales Family Lycopodiaceae or FamilyHuperziaceae. In some embodiments the nucleic acid molecule encoding aPtIP-65 polypeptide is derived from a species in the GenusAustrolycopodium, Dendrolycopodium, Diphasiastrum, Diphasium, Huperzia,Lateristachys, Lycopodiastrum, Lycopodiella, Lycopodium, Palhinhaea,Pseudodiphasium, Pseudolycopodiella, Pseudolycopodium or Spinulum.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the Genus Lycopodium. In someembodiments the nucleic acid molecule encoding a PtIP-65 polypeptide isderived from a Lycopodium species selected from but not limited toLycopodium alpinum L., Lycopodium annotinum L., Lycopodium clavatum L.,Lycopodium complanatum L., Lycopodium dendroideum Michx., Lycopodiumdigitatum, Lycopodium xhabereri, Lycopodium hickeyi, Lycopodium×issleri,Lycopodium lagopus, Lycopodium obscurum L., Lycopodium phlegmaria L.,Lycopodium sabinifolium, Lycopodium sitchense, Lycopodium tristachyum,Lycopodium venustulum, Lycopodium venustulum var. venustulum, Lycopodiumvenustulum var. verticale, Lycopodium volubile and Lycopodium×zeilleri.

In some embodiments the nucleic acid molecule encoding a PtIP-65polypeptide is derived from a species in the Genus Huperzia. In someembodiments the nucleic acid molecule encoding a PtIP-65 polypeptide isderived from a species selected from but not limited to Huperziaappressa, Huperzia arctica, Huperzia attenuata, Huperzia australiana,Huperzia balansae, Huperzia billardierei, Huperzia brassii, Huperziacampiana, Huperzia capellae, Huperzia carinata, Huperzia cf. carinataARF000603, Huperzia cf. nummulariifolia ARF001140, Huperzia cf.phlegmaria ARF000717, Huperzia cf. phlegmaria ARF000771, Huperzia cf.phlegmaria ARF000785, Huperzia cf. phlegmaria ARF001007, Huperzia cf.phlegmaria ARF002568, Huperzia cf. phlegmaria ARF002703, Huperzia cf.phlegmaria Wikstrom 1998, Huperzia chinensis, Huperzia compacta,Huperzia crassa, Huperzia crispata, Huperzia cryptomeriana, Huperziacumingii, Huperzia dacrydioides, Huperzia dalhousieana, Huperziadichotoma, Huperzia emeiensis, Huperzia ericifolia, Huperzia eversa,Huperzia fargesii, Huperzia fordii, Huperzia funiformis, Huperziagoebellii, Huperzia haleakalae, Huperzia hamiltonii, Huperziaheteroclita, Huperzia hippuridea, Huperzia hippuris, Huperzia holstii,Huperzia horizontalis, Huperzia hunanensis, Huperzia hystrix, Huperzialindenii, Huperzia linifolia, Huperzia lockyeri, Huperzia lucidula,Huperzia mingcheensis, Huperzia miyoshiana, Huperzia nanchuanensis,Huperzia nummulariifolia, Huperzia obtusifolia, Huperziaophioglossoides, Huperzia petiolate, Huperzia phlegmaria, Huperziaphlegmarioides, Huperzia phyllantha, Huperzia pinifolia, Huperziapolydactyla, Huperzia prolifera, Huperzia reflexa, Huperziarosenstockiana, Huperzia rufescens, Huperzia salvinoides, Huperziasarmentosa, Huperzia selago, Huperzia serrata, Huperzia sieboldii,Huperzia somae, Huperzia squarrosa, Huperzia subulata, Huperziasutchueniana, Huperzia tauri, Huperzia taxifolia, Huperzia tenuis,Huperzia tetragona, Huperzia tetrasticha, Huperzia unguiculata, Huperziavaria, Huperzia verticillata and Huperzia wilsonii.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); and at least one amino acid sequence motif selected from:i) an amino acid sequence motif 2 as represented by an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW]RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); and ii) an amino acid sequence motif 3 as representedby an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); and at least one amino acid sequence motif selectedfrom: i) an amino acid sequence motif 1 as represented by an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); ii) an amino acid sequence motif 5 as represented byan amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); iii) an amino acid sequence motif 12 as represented byan amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS](SEQ ID NO: 158); andiv) an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO:162).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); ii) an amino acid sequence motif 17 as represented byan amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); iii) anamino acid sequence motif 20 as represented by an amino acid sequence ofthe formula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO:166); iv) an amino acid sequence motif 24 as represented by an aminoacid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and v) an amino acid sequence motif 26 as representedby an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); anamino acid sequence motif 11 as represented by an amino acid sequence ofthe formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); an aminoacid sequence motif 20 as represented by an amino acid sequence of theformula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166);an amino acid sequence motif 24 as represented by an amino acid sequenceof the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and an amino acid sequence motif 26 as represented byan amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide an amino acid sequence motif 12 asrepresented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 as represented by an amino acid sequence ofthe formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ IDNO: 152); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 11 as represented by anamino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 20 as represented by anamino acid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF](SEQ ID NO: 166); an aminoacid sequence motif 26 as represented by an amino acid sequence of theformula L[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQID NO: 171) and an amino acid sequence motif 24 as represented by anamino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 as represented by an amino acid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); ii) an amino acid sequence motif 13 as represented by an aminoacid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO:159); iii) an amino acid sequence motif 19 as represented by an aminoacid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); iv)an amino acid sequence motif 22 as represented by an amino acid sequenceof the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); andv) an amino acid sequence motif 23 as represented by an amino acidsequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); anamino acid sequence motif 8 as represented by an amino acid sequence ofthe formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); an amino acid sequence motif 13 as represented by an aminoacid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO:159); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 22 as represented by an amino acid sequence ofthe formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); an aminoacid sequence motif 23 as represented by an amino acid sequence of theformula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169); and at least one amino acidsequence motif selected from: i) an amino acid sequence motif 4 asrepresented by an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); ii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iii) an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide, an amino acid sequence motif 12as represented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 as represented by an amino acid sequence ofthe formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 as represented by an amino acid sequence ofthe formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif selected from: an amino acid sequence motif 1 asrepresented by an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 as represented by an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 23 as represented by anamino acid sequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169);an amino acid sequence motif 4 as represented by an amino acid sequenceof the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ IDNO: 159); an amino acid sequence motif 22 as represented by an aminoacid sequence of the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ IDNO: 168); and an amino acid sequence motif 8 as represented by an aminoacid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 2 asrepresented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 as represented by an amino acid sequence ofthe formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 10 as represented by an amino acid sequence of theformula[WPY][DTN][HNV][VI][AG]R[VI][RW]F[SQ]TS[RL][GL]R[TI]FE[YF]G[SM][ST][RV][EQ]P[ST]G[QKR][WV][FIT][TE][VA]NVGSGVCVG[VM](SEQ ID NO: 156); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 27 as represented by anamino acid sequence of the formula T[TFPS]IL[EQS][EQH]LP[DA] (SEQ ID NO:172); an amino acid sequence motif 7 as represented by an amino acidsequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 21 as represented by anamino acid sequence of the formulaHL[TV][GA][NRS][AND]I[STL][IVL][ND][VILM][DTS]LGI[ND][TA]P[TK][VI]V[PA](SEQ ID NO: 167); an amino acid sequence motif 18 as represented by anamino acid sequence of the formulaG[TLP][AE]GAST[NTA]F[GQH]W[EQ]T[VA]R[TA]FPS[TIS]NA (SEQ ID NO: 164); anamino acid sequence motif 9 as represented by an amino acid sequence ofthe formula[IMV][QE]G[ST]I[AS][ND]L[TI]V[SRT]T[NAD][EA]YSVW[CG]H[IV]SDT[LVI]APAQ[SL][LI][PI][KS][HR][RASV]A (SEQ ID NO: 155); an amino acid sequence motif 14 asrepresented by an amino acid sequence of the formula[LW][VI]GEG[RS]I[TD][AN]L[PQ]CSA[NR]IQV[FIL]T[SD][GAS][GAD][NGY][NDG][LF]PF[AG][TS]F[ST]FPV[RGQ][LSV][LF]Y[DNS][GA][GR]AHS[DNQT]VQ[VI][LI](SEQ ID NO: 160).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQID NO: 152); an amino acid sequence motif 2 as represented by an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 as represented by an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163);ii) an amino acid sequence motif 4 as represented by an amino acidsequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); iii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iv) an amino acid sequence motif 15 as representedby an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12as represented by an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 2 as represented by an amino acid sequenceof the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ IDNO: 152); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 as represented by an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); and an amino acid sequence motif 4 as represented byan amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); and at least one aminoacid sequence motif selected from: i) an amino acid sequence motif 2having at least 90% identity to an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); and ii) an amino acid sequence motif 3 having at least90% identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); and at least one amino acid sequence motif selectedfrom: i) an amino acid sequence motif 1 having at least 90% identity toan amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); ii) an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); iii) an amino acid sequence motif 12 having at least90% identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); andiv) an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 11 having at least 90% identity to an aminoacid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); ii) an amino acid sequence motif 17 having at least90% identity to an amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); iii) anamino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166); iv) anamino acid sequence motif 24 having at least 90% identity to an aminoacid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and v) an amino acid sequence motif 26 having at least90% identity to an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); an amino acid sequence motif 11 having at least 90% identityto an amino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 17 having at least 90%identity to an amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); an aminoacid sequence motif 20 having at least 90% identity to an amino acidsequence of the formula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF](SEQ ID NO: 166); an amino acid sequence motif 24 having at least 90%identity to an amino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and an amino acid sequence motif 26 having at least90% identity to an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 17 having at least 90% identity to an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); anamino acid sequence motif selected from: an amino acid sequence motif 1having at least 90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 11 having at least 90%identity to an amino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 20 having at least 90%identity to an amino acid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166); an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171) and an amino acid sequence motif 24 having at least 90% identity toan amino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 8 having at least 90% identity to an aminoacid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); ii) an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO: 159); iii) an amino acidsequence motif 19 having at least 90% identity to an amino acid sequenceof the formula [TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQID NO: 165); iv) an amino acid sequence motif 22 having at least 90%identity to an amino acid sequence of the formula[KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); and v) an amino acidsequence motif 23 having at least 90% identity to an amino acid sequenceof the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); an amino acid sequence motif 8 having at least 90% identity toan amino acid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); an amino acid sequence motif 13 having at least 90% identityto an amino acid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD](SEQ ID NO: 159); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 22 having at least 90% identity to an aminoacid sequence of the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ IDNO: 168); an amino acid sequence motif 23 having at least 90% identityto an amino acid sequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO:169); and at least one amino acid sequence motif selected from: i) anamino acid sequence motif 4 having at least 90% identity to an aminoacid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); ii) an amino acid sequence motif 7 having at least 90%identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iii) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO:152); an amino acid sequence motif selected from: an amino acid sequencemotif 1 having at least 90% identity to an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 having at least 90% identity to an amino acid sequenceof the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 23 having at least 90%identity to an amino acid sequence of the formula Y[TA][ST]TQEESRTLT(SEQ ID NO: 169); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO: 159); an amino acid sequencemotif 22 having at least 90% identity to an amino acid sequence of theformula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); and an aminoacid sequence motif 8 having at least 90% identity to an amino acidsequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 2having at least 90% identity to an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO:152); an amino acid sequence motif 10 having at least 90% identity to anamino acid sequence of the formula[WPY][DTN][HNV][VI][AG]R[VI][RW]F[SQ]TS[RL][GL]R[TI]FE[YF]G[SM][ST][RV][EQ]P[ST]G[QKR][WV][FIT][TE][VA]NVGSGVCVG[VM](SEQ ID NO: 156); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 27 having at least 90%identity to an amino acid sequence of the formulaT[TFPS]IL[EQS][EQH]LP[DA] (SEQ ID NO: 172); an amino acid sequence motif7 having at least 90% identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 21 having at least 90%identity to an amino acid sequence of the formulaHL[TV][GA][NRS][AND]I[STL][IVL][ND][VILM][DTS]LGI[ND][TA]P[TK][VI]V[PA](SEQ ID NO: 167); an amino acid sequence motif 18 having at least 90%identity to an amino acid sequence of the formulaG[TLP][AE]GAST[NTA]F[GQH]W[EQ]T[VA]R[TA]FPS[TIS]NA (SEQ ID NO: 164); anamino acid sequence motif 9 having at least 90% identity to an aminoacid sequence of the formula[IMV][QE]G[ST]I[AS][ND]L[TI]V[SRT]T[NAD][EA]YSVW[CG]H[IV]SDT[LVI]APAQ[SL][LI][PI][KS][HR][RASV]A (SEQ ID NO: 155v) an amino acid sequence motif 14 havingat least 90% identity to an amino acid sequence of the formula[LW][VI]GEG[RS]I[TD][AN]L[PQ]CSA[NR]IQV[FIL]T[SD][GAS][GAD][NGY][NDG][LF]PF[AG][TS]F[ST]FPV[RGQ][LSV][LF]Y[DNS][GA][GR]AHS[DNQT]VQ[VI][LI](SEQ ID NO: 160).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 having at least 90% identity to an amino acid sequence of theformulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); ii) an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); iii) an amino acid sequence motif 7 having at least90% identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iv) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 17 having at least 90% identity to an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); anamino acid sequence motif selected from: an amino acid sequence motif 1having at least 90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 having at least 90% identity to an amino acid sequenceof the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); and an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); and at least oneamino acid sequence motif selected from: i) an amino acid sequence motif2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); and ii) an amino acid sequence motif 3 as representedby an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); and at least one amino acid sequencemotif selected from: i) an amino acid sequence motif 1 as represented byan amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); ii) an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); iii) anamino acid sequence motif 12 as represented by an amino acid sequence ofthe formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); and iv) an amino acid sequence motif 16 as representedby an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); ii) anamino acid sequence motif 17 as represented by an amino acid sequence ofthe formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); iii) an amino acid sequence motif 20 as represented byan amino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); iv) an amino acid sequence motif 24 as represented by an aminoacid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and v) an aminoacid sequence motif 26 as represented by an amino acid sequence of theformulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 17 as represented by an amino acid sequence of theformula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ IDNO: 189); an amino acid sequence motif 20 as represented by an aminoacid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 24 as represented by an amino acidsequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and an aminoacid sequence motif 26 as represented by an amino acid sequence of theformulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: at least one amino acid sequencemotif selected from: an amino acid sequence motif 12 as represented byan amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 11 asrepresented by an amino acid sequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 5 as represented by an amino acid sequence of theformula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 20 as represented by an amino acid sequence of theformula DT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQID NO: 192); an amino acid sequence motif 26 as represented by an aminoacid sequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197) and an amino acid sequence motif 24 as represented byan amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 as represented by an amino acid sequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); ii) an amino acid sequence motif 13 as represented byan amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); iii) an aminoacid sequence motif 19 as represented by an amino acid sequence of theformula [TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL](SEQ ID NO: 191); iv) an amino acid sequence motif 22 as represented byan amino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and v)an amino acid sequence motif 23 as represented by an amino acid sequenceof the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 8 as represented by an amino acid sequence of theformula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 22 as represented by an amino acidsequence of the formula [KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ(SEQ ID NO: 194); an amino acid sequence motif 23 as represented by anamino acid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195);and at least one amino acid sequence motif selected from: i) an aminoacid sequence motif 4 as represented by an amino acid sequence of theformulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); ii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iii) an amino acid sequence motif as representedby an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: at least one amino acid sequencemotif selected from: an amino acid sequence motif 12 as represented byan amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 as represented by an amino acidsequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO:178); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 as represented by an amino acid sequence ofthe formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177) an aminoacid sequence motif 23 as represented by an amino acid sequence of theformula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); an amino acid sequencemotif 4 as represented by an amino acid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acid sequence motif 22 as represented by anamino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and anamino acid sequence motif 8 as represented by an amino acid sequence ofthe formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: at least one amino acid sequencemotif selected from: an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 as represented by an amino acidsequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO:178); an amino acid sequence motif 10 as represented by an amino acidsequence of the formula[WPYF][DTNQES][HNVQIL][VIL][AG]R[VIL][RWKYH]F[SQTN]TS[RLKIV][GLIV]R[TISLV]FE[YFW]G[SMT][ST][RV][EQND]P[ST]G[QKRN][WVYIL][FITLVS][TESD][VAIL]NVGSGVCVG[VMIL] (SEQ ID NO: 182); an amino acid sequence motif 3 as represented byan amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 27 asrepresented by an amino acid sequence of the formulaT[TFPS]IL[EQSDNT][EQHDN]LP[DAE] (SEQ ID NO: 198); an amino acid sequencemotif 7 as represented by an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 21 as represented by anamino acid sequence of the formulaHL[TVSIL][GA][NRSQKT][ANDQE]I[STLIV][IVL][NDQE][VILM][DTSE]LGI[ND][TAS]P[TKSR][VIL]V[PA] (SEQ ID NO: 193); an amino acid sequencemotif 18 as represented by an amino acid sequence of the formulaG[TLPSIV][AED]GAST[NTAQS]F[GQHN]W[EQND]T[VAIL]R[TAS]FPS[TISLV]NA (SEQ IDNO: 190); an amino acid sequence motif 9 as represented by an amino acidsequence of the formula[IMVL][QEND]G[ST]I[AST][NDQE]L[TISLV]V[SRTK]T[NADQE][EAD]YSVW[CG]H[IVL]SDT[LVI]APAQ[SLTIV][LIV][PIVL][KSRT][HRK][RASVKTIL]A (SEQ ID NO: 181); anamino acid sequence motif 14 as represented by an amino acid sequence ofthe formula[LWIVF][VIL]GEG[RSKT]I[TDSE][ANQ]L[PQN]CSA[NRQK]IQV[FILV]T[SDTE][GAST][GADE][NGYQ][NDGQE][LFIV]PF[AG][TS]F[ST]FPV[RGQKN][LSVIT][LFIV]Y[DNSQET][GA][GRK]AHS[DNQTES]VQ[VIL][LIV] (SEQ ID NO: 186).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 as represented byan amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 as represented by an amino acid sequence of theformula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ IDNO: 189); ii) an amino acid sequence motif 4 as represented by an aminoacid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); iii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iv) an amino acid sequence motif 15 as representedby an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: at least one amino acid sequencemotif selected from: an amino acid sequence motif 12 as represented byan amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 as represented by an amino acid sequence ofthe formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); and anamino acid sequence motif 4 as represented by an amino acid sequence ofthe formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising, an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide comprising, an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); and at least oneamino acid sequence motif selected from: i) an amino acid sequence motif2 having at least 90% identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); and ii) an amino acid sequence motif 3 having at least90% identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); and at least one amino acid sequencemotif selected from: i) an amino acid sequence motif 1 having at least90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); ii) an amino acid sequence motif 5 having at least 90% identity toan amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); iii) anamino acid sequence motif 12 having at least 90% identity to an aminoacid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); and iv) an amino acid sequence motif 16 having atleast 90% identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 having at least 90% identity to an amino acid sequenceof the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); ii) anamino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); iii) an amino acid sequence motif 20 having at least 90% identityto an amino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV](SEQ ID NO:192); iv) an amino acid sequence motif 24 having at least 90% identityto an amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and v) an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 11 having at least 90% identity to an amino acidsequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 17 having at least 90% identity to an amino acidsequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif 20 having at least 90% identity to anamino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 24 having at least 90% identity to anamino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQID NO: 184); an amino acid sequence motif 2 having at least 90% identityto an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 11having at least 90% identity to an amino acid sequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 5 having at least 90% identity to an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 20 having at least 90% identity to an amino acidsequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 26 having at least 90% identity to anamino acid sequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197) and an amino acid sequence motif 24 having at least 90%identity to an amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 having at least 90% identity to an amino acid sequenceof the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); ii) an amino acid sequence motif 13 having at least90% identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); iii) an aminoacid sequence motif 19 having at least 90% identity to an amino acidsequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); iv) an amino acid sequence motif 22 having at least 90% identityto an amino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and v)an amino acid sequence motif 23 having at least 90% identity to an aminoacid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 8 having at least 90% identity to an amino acidsequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acidsequence motif 19 having at least 90% identity to an amino acid sequenceof the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 22 having at least 90% identity to anamino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); anamino acid sequence motif 23 having at least 90% identity to an aminoacid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); andat least one amino acid sequence motif selected from: i) an amino acidsequence motif 4 having at least 90% identity to an amino acid sequenceof the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); ii) an amino acid sequence motif 7 having at least 90%identity to an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iii) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQID NO: 184); an amino acid sequence motif 2 having at least 90% identityto an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 having at least 90% identity to anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15having at least 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 having at least 90% identity to an aminoacid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 23 having at least 90% identity to an amino acidsequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); an aminoacid sequence motif 4 having at least 90% identity to an amino acidsequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); an amino acidsequence motif 22 having at least 90% identity to an amino acid sequenceof the formula [KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ IDNO: 194); and an amino acid sequence motif 8 having at least 90%identity to an amino acid sequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 2having at least 90% identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 having at least 90% identity to anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 10 having at least 90%identity to an amino acid sequence of the formula[WPYF][DTNQES][HNVQIL][VIL][AG]R[VIL][RWKYH]F[SQTN]TS[RLKIV][GLIV]R[TISLV]FE[YFW]G[SMT][ST][RV][EQND]P[ST]G[QKRN][WVYIL][FITLVS][TESD][VAIL]NVGSGVCVG[VMIL] (SEQ ID NO: 182); an amino acid sequence motif 3 having at least90% identity to an amino acid sequence ofGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 27having at least 90% identity to an amino acid sequence of the formulaT[TFPS]IL[EQSDNT][EQHDN]LP[DAE] (SEQ ID NO: 198); an amino acid sequencemotif 7 having at least 90% identity to an amino acid sequence of theformula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 21 having at least 90%identity to an amino acid sequence of the formulaHL[TVSIL][GA][NRSQKT][ANDQE]I[STLIV][IVL][NDQE][VILM][DTSE]LGI[ND][TAS]P[TKSR][VIL]V[PA] (SEQ ID NO: 193); an amino acid sequencemotif 18 having at least 90% identity to an amino acid sequence of theformula G[TLPSIV][AED]GAST[NTAQS]F[GQHN]W[EQND]T[VAIL]R[TAS]FPS[TISLV]NA(SEQ ID NO: 190); an amino acid sequence motif 9 having at least 90%identity to an amino acid sequence of the formula[IMVL][QEND]G[ST]I[AST][NDQE]L[TISLV]V[SRTK]T[NADQE][EAD]YSVW[CG]H[IVL]SDT[LVI]APAQ[SLTIV][LIV][PIVL][KSRT][HRK][RASVKTIL]A (SEQ ID NO: 181); anamino acid sequence motif 14 having at least 90% identity to an aminoacid sequence of the formula[LWIVF][VIL]GEG[RSKT]I[TDSE][ANQ]L[PQN]CSA[NRQK]IQV[FILV]T[SDTE][GAST][GADE][NGYQ][NDGQE][LFIV]PF[AG][TS]F[ST]FPV[RGQKN][LSVIT][LFIV]Y[DNSQET][GA][GRK]AHS[DNQTES]VQ[VIL][LIV] (SEQ ID NO: 186).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising an amino acid sequence motif 6 having at least90% identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence ofGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); ii) an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); iii) an amino acid sequence motif 7 having at least90% identity to an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iv) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187). Insome embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus of the PtIP-65 polypeptide: an amino acid sequence motif 12having at least 90% identity to an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15having at least 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 having at least 90% identity to an aminoacid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); and anamino acid sequence motif 4 having at least 90% identity to an aminoacid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176).

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide that is about 300 to 400 amino acids in length having aconsensus secondary structure comprising 13 to 17 segments ofpredominately beta strand structure separated by coil regions.

In some embodiments the nucleic acid molecule encodes a PtIP-65polypeptide, comprising, sequentially from the N-terminus to theC-terminus; i) a beta strand-1 (β1) of between about 3 amino acids andabout 9 amino acids in length, ii) a coil of between about 17 aminoacids and about 21 amino acids in length; iii) a beta strand-2 (β2) ofbetween about 7 amino acids and about 9 amino acids in length, iv) acoil of between about 3 amino acids and about 4 amino acids in length;v) a beta strand-3 (β3) of between about 9 amino acids and about 10amino acids in length, vi) a coil of between about 2 amino acids andabout 5 amino acids in length; vii) a beta strand-4 (β4) of betweenabout 3 amino acids and about 8 amino acids in length, viii) a coil ofbetween about 7 amino acids and about 13 amino acids in length; ix) abeta strand-5 (β5) of between about 3 amino acids and about 5 aminoacids in length, x) a coil of between about 4 amino acids and about 5amino acids in length; xi) a beta strand-6 (β6) of between about 8 aminoacids and about 12 amino acids in length, xii) a coil of between about 5amino acids and about 12 amino acids in length; xiii) a beta strand-7(β7) of between about 9 amino acids and about 13 amino acids in length,xiv) a coil of about 4 amino acids in length; xv) a beta strand-8 (β1)of between about 3 amino acids and about 7 amino acids in length, xvi) acoil of between about 5 amino acids and about 9 amino acids in length;xvii) a beta strand-9 (β9) of between about 3 amino acids and about 5amino acids in length, xviii) a coil of between about 3 amino acids andabout 6 amino acids in length; ixx) a beta strand-10 (β10) of betweenabout 7 amino acids and about 13 amino acids in length, xx) a coil ofbetween about 3 amino acids and about 8 amino acids in length; xxi) abeta strand-11 (β11) of between about 5 amino acids and about 18 aminoacids in length, xxii) a coil of between about 10 amino acids and about22 amino acids in length; xxiii) a beta strand-12 (β12) of between about7 amino acids and about 12 amino acids in length, xxiv) a coil ofbetween about 5 amino acids and about 13 amino acids in length; xxv) abeta strand-13 (β13) of between about 60 amino acids and about 90 aminoacids in length, xxvi) a coil of between about 4 amino acids and about 8amino acids in length; xxvii) a beta strand-14 (β14) of between about 22amino acids and about 30 amino acids in length, xxviii) a coil ofbetween about 3 amino acids and about 8 amino acids in length; and xxix)a beta strand-15 (β15) of between about 22 amino acids and about 26amino acids in length. As used herein, the term “about” when used in thecontext of the lower/upper limit of the length of a secondary structuralelement means the greater of −/+ an integer of up to −/+20% of thelength of the secondary structural element or −/+1 amino acid. By meansof example, a secondary structure element of between about 3 amino acidsand about 23 amino acids in length means a secondary structure elementof between 2 and 27 amino acids in length.

Also provided are nucleic acid molecules that encode transcriptionand/or translation products that are subsequently spliced to ultimatelyproduce functional PtIP-50 polypeptides or PtIP-65 polypeptides.Splicing can be accomplished in vitro or in vivo, and can involve cis-or trans-splicing. The substrate for splicing can be polynucleotides(e.g., RNA transcripts) or polypeptides. An example of cis-splicing of apolynucleotide is where an intron inserted into a coding sequence isremoved and the two flanking exon regions are spliced to generate aPtIP-50 polypeptide or PtIP-65 polypeptide encoding sequence. An exampleof trans splicing would be where a polynucleotide is encrypted byseparating the coding sequence into two or more fragments that can beseparately transcribed and then spliced to form the full-lengthpesticidal encoding sequence. The use of a splicing enhancer sequence,which can be introduced into a construct, can facilitate splicing eitherin cis or trans-splicing of polypeptides (U.S. Pat. Nos. 6,365,377 and6,531,316). Thus, in some embodiments the polynucleotides do notdirectly encode a full-length PtIP-50 polypeptide or PtIP-65polypeptide, but rather encode a fragment or fragments of a PtIP-50polypeptide or PtIP-65 polypeptide. These polynucleotides can be used toexpress a functional PtIP-50 polypeptide or PtIP-65 polypeptide througha mechanism involving splicing, where splicing can occur at the level ofpolynucleotide (e.g., intron/exon) and/or polypeptide (e.g.,intein/extein). This can be useful, for example, in controllingexpression of pesticidal activity, since a functional pesticidalpolypeptide will only be expressed if all required fragments areexpressed in an environment that permits splicing processes to generatefunctional product. In another example, introduction of one or moreinsertion sequences into a polynucleotide can facilitate recombinationwith a low homology polynucleotide; use of an intron or intein for theinsertion sequence facilitates the removal of the intervening sequence,thereby restoring function of the encoded variant.

Nucleic acid molecules that are fragments of these nucleic acidsequences encoding PtIP-50 polypeptides or PtIP-65 polypeptides are alsoencompassed by the embodiments. “Fragment” as used herein refers to aportion of the nucleic acid sequence encoding a PtIP-50 polypeptide orPtIP-65 polypeptide. A fragment of a nucleic acid sequence may encode abiologically active portion of a PtIP-50 polypeptide or PtIP-65polypeptide or it may be a fragment that can be used as a hybridizationprobe or PCR primer using methods disclosed below. Nucleic acidmolecules that are fragments of a nucleic acid sequence encoding aPtIP-50 polypeptide or PtIP-65 polypeptide comprise at least about 150,180, 210, 240, 270, 300, 330 or 360, contiguous nucleotides or up to thenumber of nucleotides present in a full-length nucleic acid sequenceencoding a PtIP-50 polypeptide or PtIP-65 polypeptide disclosed herein,depending upon the intended use. “Contiguous nucleotides” is used hereinto refer to nucleotide residues that are immediately adjacent to oneanother. Fragments of the nucleic acid sequences of the embodiments willencode protein fragments that retain the biological activity of thePtIP-50 polypeptide/PtIP-65 polypeptide and, hence, retain insecticidalactivity. “Retains insecticidal activity” is used herein to refer to apolypeptide having at least about 10%, at least about 30%, at leastabout 50%, at least about 70%, 80%, 90%, 95% or higher of theinsecticidal activity of the full-length PtIP-50Aa polypeptide (SEQ IDNO: 71) in combination with PtIP-65Aa polypeptide (SEQ ID NO: 22). Insome embodiments, the insecticidal activity is Lepidoptera activity. Inone embodiment, the insecticidal activity is against a Coleopteranspecies. In some embodiments, the insecticidal activity is against oneor more insect pests of the corn rootworm complex: western cornrootworm, Diabrotica virgifera; northern corn rootworm, D. barberi:Southern corn rootworm or spotted cucumber beetle; Diabroticaundecimpunctata howardi, and the Mexican corn rootworm, D. virgiferazeae. In one embodiment, the insecticidal activity is against aDiabrotica species.

In some embodiments a fragment of a nucleic acid sequence encoding aPtIP-50 polypeptide encoding a biologically active portion of a proteinwill encode at least about 15, 20, 30, 50, 75, 100, 125, contiguousamino acids or up to the total number of amino acids present in afull-length PtIP-50 polypeptide of the embodiments. In some embodiments,the fragment is an N-terminal and/or a C-terminal truncation of at leastabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more aminoacids from the N-terminus and/or C-terminus relative to SEQ ID NO: 71,SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ IDNO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90,SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98 or variants thereof,e.g., by proteolysis, insertion of a start codon, deletion of the codonsencoding the deleted amino acids with the concomitant insertion of astop codon or by insertion of a stop codon in the coding sequence. Insome embodiments, the fragments encompassed herein result from theremoval of the N-terminal 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more amino acids from theN-terminus relative to SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO:83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ IDNO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 orSEQ ID NO: 98 or variants thereof, e.g., by proteolysis or by insertionof a start codon in the coding sequence. In some embodiments, thefragments encompassed herein result from the removal of the N-terminal1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 amino acids relative toSEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO:75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ IDNO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89,SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO:94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98 orvariants thereof, e.g., by proteolysis or by insertion of a start codonin the coding sequence.

In some embodiments a PtIP-50 polypeptide is encoded by a nucleic acidsequence sufficiently homologous to the nucleic acid sequence of SEQ IDNO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52,SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ IDNO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ ID NO: 70. “Sufficientlyhomologous” is used herein to refer to an amino acid or nucleic acidsequence that has at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or greater sequence homology compared to a referencesequence using one of the alignment programs described herein usingstandard parameters. One of skill in the art will recognize that thesevalues can be appropriately adjusted to determine corresponding homologyof proteins encoded by two nucleic acid sequences by taking into accountcodon degeneracy, amino acid similarity, reading frame positioning, andthe like. In some embodiments the sequence homology is against the fulllength sequence of the polynucleotide encoding a PtIP-50 polypeptide oragainst the full length sequence of a PtIP-50 polypeptide. In someembodiments the PtIP-50 polypeptide has at least about 50%, 55%, 60%,65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identitycompared to SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO:98. In some embodiments the sequence identity is calculated usingClustalW algorithm in the ALIGNX® module of the Vector NTI® ProgramSuite (Invitrogen Corporation, Carlsbad, Calif.) with all defaultparameters. In some embodiments the sequence identity is across theentire length of polypeptide calculated using ClustalW algorithm in theALIGNX module of the Vector NTI Program Suite (Invitrogen Corporation,Carlsbad, Calif.) with all default parameters.

To determine the percent identity of two amino acid sequences or of twonucleic acid sequences, the sequences are aligned for optimal comparisonpurposes. The percent identity between the two sequences is a functionof the number of identical positions shared by the sequences (i.e.,percent identity=number of identical positions/total number of positions(e.g., overlapping positions)×100). In one embodiment, the two sequencesare the same length. In another embodiment, the comparison is across theentirety of the reference sequence (e.g., across the entirety of SEQ IDNO: 43). The percent identity between two sequences can be determinedusing techniques similar to those described below, with or withoutallowing gaps. In calculating percent identity, typically exact matchesare counted.

The determination of percent identity between two sequences can beaccomplished using a mathematical algorithm. A non-limiting example of amathematical algorithm utilized for the comparison of two sequences isthe algorithm of Karlin and Altschul, (1990) Proc. Natl. Acad. Sci. USA87:2264, modified as in Karlin and Altschul, (1993) Proc. Natl. Acad.Sci. USA 90:5873-5877. Such an algorithm is incorporated into the BLASTNand BLASTX programs of Altschul, et al., (1990) J. Mol. Biol. 215:403.BLAST nucleotide searches can be performed with the BLASTN program,score=100, wordlength=12, to obtain nucleic acid sequences homologous topesticidal nucleic acid molecules of the embodiments. BLAST proteinsearches can be performed with the BLASTX program, score=50,wordlength=3, to obtain amino acid sequences homologous to pesticidalprotein molecules of the embodiments. To obtain gapped alignments forcomparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized asdescribed in Altschul, et al., (1997) Nucleic Acids Res. 25:3389.Alternatively, PSI-Blast can be used to perform an iterated search thatdetects distant relationships between molecules. See, Altschul, et al.,(1997) supra. When utilizing BLAST, Gapped BLAST, and PSI-Blastprograms, the default parameters of the respective programs (e.g.,BLASTX and BLASTN) can be used. Alignment may also be performed manuallyby inspection.

Another non-limiting example of a mathematical algorithm utilized forthe comparison of sequences is the ClustalW algorithm (Higgins, et al.,(1994) Nucleic Acids Res. 22:4673-4680). ClustalW compares sequences andaligns the entirety of the amino acid or DNA sequence, and thus canprovide data about the sequence conservation of the entire amino acidsequence. The ClustalW algorithm is used in several commerciallyavailable DNA/amino acid analysis software packages, such as the ALIGNX®module of the Vector NTI® Program Suite (Invitrogen Corporation,Carlsbad, Calif.). After alignment of amino acid sequences withClustalW, the percent amino acid identity can be assessed. Anon-limiting example of a software program useful for analysis ofClustalW alignments is GENEDOC™ GENEDOC™ (Karl Nicholas) allowsassessment of amino acid (or DNA) similarity and identity betweenmultiple proteins. Another non-limiting example of a mathematicalalgorithm utilized for the comparison of sequences is the algorithm ofMyers and Miller, (1988) CABIOS 4:11-17. Such an algorithm isincorporated into the ALIGN program (version 2.0), which is part of theGCG Wisconsin Genetics Software Package, Version 10 (available fromAccelrys, Inc., 9685 Scranton Rd., San Diego, Calif., USA). Whenutilizing the ALIGN program for comparing amino acid sequences, a PAM120weight residue table, a gap length penalty of 12, and a gap penalty of 4can be used.

Another non-limiting example of a mathematical algorithm utilized forthe comparison of sequences is the algorithm of Needleman and Wunsch,(1970) J. Mol. Biol. 48(3):443-453, used GAP Version 10 software todetermine sequence identity or similarity using the following defaultparameters: % identity and % similarity for a nucleic acid sequenceusing GAP Weight of 50 and Length Weight of 3, and the nwsgapdna.cmpiiscoring matrix; % identity or % similarity for an amino acid sequenceusing GAP weight of 8 and length weight of 2, and the BLOSUM62 scoringprogram. Equivalent programs may also be used. “Equivalent program” isused herein to refer to any sequence comparison program that, for anytwo sequences in question, generates an alignment having identicalnucleotide residue matches and an identical percent sequence identitywhen compared to the corresponding alignment generated by GAP Version10.

The embodiments also encompass nucleic acid molecules encoding PtIP-50polypeptide variants. “Variants” of the PtIP-50 polypeptide encodingnucleic acid sequences include those sequences that encode the PtIP-50polypeptides disclosed herein but that differ conservatively because ofthe degeneracy of the genetic code as well as those that aresufficiently identical as discussed above. Naturally occurring allelicvariants can be identified with the use of well-known molecular biologytechniques, such as polymerase chain reaction (PCR) and hybridizationtechniques as outlined below. Variant nucleic acid sequences alsoinclude synthetically derived nucleic acid sequences that have beengenerated, for example, by using site-directed mutagenesis but whichstill encode the PtIP-50 polypeptides disclosed as discussed below.

The present disclosure provides isolated or recombinant polynucleotidesthat encode any of the PtIP-50 polypeptides disclosed herein. Thosehaving ordinary skill in the art will readily appreciate that due to thedegeneracy of the genetic code, a multitude of nucleotide sequencesencoding PtIP-50 polypeptides of the present disclosure exist. Table 1is a codon table that provides the synonymous codons for each aminoacid. For example, the codons AGA, AGG, CGA, CGC, CGG, and CGU allencode the amino acid arginine. Thus, at every position in the nucleicacids of the disclosure where an arginine is specified by a codon, thecodon can be altered to any of the corresponding codons described abovewithout altering the encoded polypeptide. It is understood that U in anRNA sequence corresponds to T in a DNA sequence.

TABLE 1 Alanine Ala GCA GCC GCG GC Cysteine Cys UGC UGU Aspartic ac dAsp GAC GA Glutamic acid Glu GAA GAG Phenylalanine Phe UUC UUU G ycineGly GGA GGC GGG GGU His idine His CAC CAU Isoleucine II AUA AUC AUULysine Lys AAA AAG Leucine Leu UUA UUG CUA CUC CUG C U Methionine MetAUG Aspa agine As AAC AAU Pro ine Pro CCA CCC CCG CCU Glutamine GlnCAA CAG Arginin Arg AGA AGG CGA CGC CGG CGU erine SerAGC AGU UCA UCC UCG UC Threonine Thr ACA ACC ACG ACU Valine ValGUA GUC GUG UU Tryptophan Trp UGG Tyrosine Tyr UAC UAU

The skilled artisan will further appreciate that changes can beintroduced by mutation of the nucleic acid sequences thereby leading tochanges in the amino acid sequence of the encoded PtIP-50 polypeptides,without altering the biological activity of the proteins. Thus, variantnucleic acid molecules can be created by introducing one or morenucleotide substitutions, additions and/or deletions into thecorresponding nucleic acid sequence disclosed herein, such that one ormore amino acid substitutions, additions or deletions are introducedinto the encoded protein. Mutations can be introduced by standardtechniques, such as site-directed mutagenesis and PCR-mediatedmutagenesis. Such variant nucleic acid sequences are also encompassed bythe present disclosure.

Alternatively, variant nucleic acid sequences can be made by introducingmutations randomly along all or part of the coding sequence, such as bysaturation mutagenesis, and the resultant mutants can be screened forability to confer pesticidal activity to identify mutants that retainactivity. Following mutagenesis, the encoded protein can be expressedrecombinantly, and the activity of the protein can be determined usingstandard assay techniques.

The polynucleotides of the disclosure and fragments thereof areoptionally used as substrates for a variety of recombination andrecursive recombination reactions, in addition to standard cloningmethods as set forth in, e.g., Ausubel, Berger and Sambrook, i.e., toproduce additional pesticidal polypeptide homologues and fragmentsthereof with desired properties. A variety of such reactions are known,including those developed by the inventors and their co-workers. Methodsfor producing a variant of any nucleic acid listed herein comprisingrecursively recombining such polynucleotide with a second (or more)polynucleotide, thus forming a library of variant polynucleotides arealso embodiments of the disclosure, as are the libraries produced, thecells comprising the libraries and any recombinant polynucleotideproduces by such methods. Additionally, such methods optionally compriseselecting a variant polynucleotide from such libraries based onpesticidal activity, as is wherein such recursive recombination is donein vitro or in vivo.

A variety of diversity generating protocols, including nucleic acidrecursive recombination protocols are available and fully described inthe art. The procedures can be used separately, and/or in combination toproduce one or more variants of a nucleic acid or set of nucleic acids,as well as variants of encoded proteins. Individually and collectively,these procedures provide robust, widely applicable ways of generatingdiversified nucleic acids and sets of nucleic acids (including, e.g.,nucleic acid libraries) useful, e.g., for the engineering or rapidevolution of nucleic acids, proteins, pathways, cells and/or organismswith new and/or improved characteristics.

While distinctions and classifications are made in the course of theensuing discussion for clarity, it will be appreciated that thetechniques are often not mutually exclusive. Indeed, the various methodscan be used singly or in combination, in parallel or in series, toaccess diverse sequence variants.

The result of any of the diversity generating procedures describedherein can be the generation of one or more nucleic acids, which can beselected or screened for nucleic acids with or which confer desirableproperties or that encode proteins with or which confer desirableproperties. Following diversification by one or more of the methodsherein or otherwise available to one of skill, any nucleic acids thatare produced can be selected for a desired activity or property, e.g.pesticidal activity or, such activity at a desired pH, etc. This caninclude identifying any activity that can be detected, for example, inan automated or automatable format, by any of the assays in the art,see, e.g., discussion of screening of insecticidal activity, infra. Avariety of related (or even unrelated) properties can be evaluated, inserial or in parallel, at the discretion of the practitioner.

Descriptions of a variety of diversity generating procedures forgenerating modified nucleic acid sequences, e.g., those coding forpolypeptides having pesticidal activity or fragments thereof, are foundin the following publications and the references cited therein: Soong,et al., (2000) Nat Genet 25(4):436-439; Stemmer, et al., (1999) TumorTargeting 4:1-4; Ness, et al., (1999) Nat Biotechnol 17:893-896; Chang,et al., (1999) Nat Biotechnol 17:793-797; Minshull and Stemmer, (1999)Curr Opin Chem Biol 3:284-290; Christians, et al., (1999) Nat Biotechnol17:259-264; Crameri, et al., (1998) Nature 391:288-291; Crameri, et al.,(1997) Nat Biotechnol 15:436-438; Zhang, et al., (1997) PNAS USA94:4504-4509; Patten, et al., (1997) Curr Opin Biotechnol 8:724-733;Crameri, et al., (1996) Nat Med 2:100-103; Crameri, et al., (1996) NatBiotechnol 14:315-319; Gates, et al., (1996) J Mol Biol 255:373-386;Stemmer, (1996) “Sexual PCR and Assembly PCR” In: The Encyclopedia ofMolecular Biology. VCH Publishers, New York. pp. 447-457; Crameri andStemmer, (1995) BioTechniques 18:194-195; Stemmer, et al., (1995) Gene,164:49-53; Stemmer, (1995) Science 270: 1510; Stemmer, (1995)Bio/Technology 13:549-553; Stemmer, (1994) Nature 370:389-391 andStemmer, (1994) PNAS USA 91:10747-10751.

Mutational methods of generating diversity include, for example,site-directed mutagenesis (Ling, et al., (1997) Anal Biochem254(2):157-178; Dale, et al., (1996) Methods Mol Biol 57:369-374; Smith,(1985) Ann Rev Genet 19:423-462; Botstein and Shortle, (1985) Science229:1193-1201; Carter, (1986) Biochem J 237:1-7 and Kunkel, (1987) “Theefficiency of oligonucleotide directed mutagenesis” in Nucleic Acids &Molecular Biology (Eckstein and Lilley, eds., Springer Verlag, Berlin));mutagenesis using uracil containing templates (Kunkel, (1985) PNAS USA82:488-492; Kunkel, et al., (1987) Methods Enzymol 154:367-382 and Bass,et al., (1988) Science 242:240-245); oligonucleotide-directedmutagenesis (Zoller and Smith, (1983) Methods Enzymol 100:468-500;Zoller and Smith, (1987) Methods Enzymol 154:329-350 (1987); Zoller andSmith, (1982) Nucleic Acids Res 10:6487-6500), phosphorothioate-modifiedDNA mutagenesis (Taylor, et al., (1985) Nucl Acids Res 13:8749-8764;Taylor, et al., (1985) Nucl Acids Res 13:8765-8787 (1985); Nakamaye andEckstein, (1986) Nucl Acids Res 14:9679-9698; Sayers, et al., (1988)Nucl Acids Res 16:791-802 and Sayers, et al., (1988) Nucl Acids Res16:803-814); mutagenesis using gapped duplex DNA (Kramer, et al., (1984)Nucl Acids Res 12:9441-9456; Kramer and Fritz, (1987) Methods Enzymol154:350-367; Kramer, et al., (1988) Nucl Acids Res 16:7207 and Fritz, etal., (1988) Nucl Acids Res 16:6987-6999).

Additional suitable methods include point mismatch repair (Kramer, etal., (1984) Cell 38:879-887), mutagenesis using repair-deficient hoststrains (Carter, et al., (1985) Nucl Acids Res 13:4431-4443 and Carter,(1987) Methods in Enzymol 154:382-403), deletion mutagenesis(Eghtedarzadeh and Henikoff, (1986) Nucl Acids Res 14:5115),restriction-selection and restriction-purification (Wells, et al.,(1986) Phil Trans R Soc Lond A 317:415-423), mutagenesis by total genesynthesis (Nambiar, et al., (1984) Science 223:1299-1301; Sakamar andKhorana, (1988) Nucl Acids Res 14:6361-6372; Wells, et al., (1985) Gene34:315-323 and Grundstrom, et al., (1985) Nucl Acids Res 13:3305-3316),double-strand break repair (Mandecki, (1986) PNAS USA, 83:7177-7181 andArnold, (1993) Curr Opin Biotech 4:450-455). Additional details on manyof the above methods can be found in Methods Enzymol Volume 154, whichalso describes useful controls for trouble-shooting problems withvarious mutagenesis methods.

Additional details regarding various diversity generating methods can befound in the following US patents, PCT Publications and applications andEPO publications: U.S. Pat. Nos. 5,723,323, 5,763,192, 5,814,476,5,817,483, 5,824,514, 5,976,862, 5,605,793, 5,811,238, 5,830,721,5,834,252, 5,837,458, WO 1995/22625, WO 1996/33207, WO 1997/20078, WO1997/35966, WO 1999/41402, WO 1999/41383, WO 1999/41369, WO 1999/41368,EP 752008, EP 0932670, WO 1999/23107, WO 1999/21979, WO 1998/31837, WO1998/27230, WO 1998/27230, WO 2000/00632, WO 2000/09679, WO 1998/42832,WO 1999/29902, WO 1998/41653, WO 1998/41622, WO 1998/42727, WO2000/18906, WO 2000/04190, WO 2000/42561, WO 2000/42559, WO 2000/42560,WO 2001/23401 and PCT/US01/06775.

The nucleotide sequences of the embodiments can also be used to isolatecorresponding sequences from ferns or other primitive plants,particularly a Asplenium, Polypodium Adianturn, Platycerium,Nephrolepis, Ophioglossum, Colysis, Bolbitis, Blechnurn, Selaginella,Lycopodium, and Huperzia species. In this manner, methods such as PCR,hybridization, and the like can be used to identify such sequences basedon their sequence homology to the sequences set forth herein. Sequencesthat are selected based on their sequence identity to the entiresequences set forth herein or to fragments thereof are encompassed bythe embodiments. Such sequences include sequences that are orthologs ofthe disclosed sequences. The term “orthologs” refers to genes derivedfrom a common ancestral gene and which are found in different species asa result of speciation. Genes found in different species are consideredorthologs when their nucleotide sequences and/or their encoded proteinsequences share substantial identity as defined elsewhere herein.Functions of orthologs are often highly conserved among species.

In a PCR approach, oligonucleotide primers can be designed for use inPCR reactions to amplify corresponding DNA sequences from cDNA orgenomic DNA extracted from any organism of interest. Methods fordesigning PCR primers and PCR cloning are generally known in the art andare disclosed in Sambrook, et al., (1989) Molecular Cloning: ALaboratory Manual (2d ed., Cold Spring Harbor Laboratory Press,Plainview, N.Y.), hereinafter “Sambrook”. See also, Innis, et al., eds.(1990) PCR Protocols: A Guide to Methods and Applications (AcademicPress, New York); Innis and Gelfand, eds. (1995) PCR Strategies(Academic Press, New York); and Innis and Gelfand, eds. (1999) PCRMethods Manual (Academic Press, New York). Known methods of PCR include,but are not limited to, methods using paired primers, nested primers,single specific primers, degenerate primers, gene-specific primers,vector-specific primers, partially-mismatched primers, and the like.

To identify potential PtIP-50 polypeptides and/or PtIP-65 polypeptidesfrom fern or moss collections, the fern or moss cell lysates can bescreened with antibodies generated against a PtIP-50 polypeptides and/orPtIP-65 polypeptides using Western blotting and/or ELISA methods. Thistype of assays can be performed in a high throughput fashion. Positivesamples can be further analyzed by various techniques such as antibodybased protein purification and identification. Methods of generatingantibodies are well known in the art as discussed infra.

Alternatively, mass spectrometry based protein identification method canbe used to identify homologs of PtIP-50 polypeptides and/or PtIP-65polypeptides using protocols in the literatures (Scott Patterson,(1998), 10.22, 1-24, Current Protocol in Molecular Biology published byJohn Wiley & Son Inc). Specifically, LC-MS/MS based proteinidentification method is used to associate the MS data of given celllysate or desired molecular weight enriched samples (excised fromSDS-PAGE gel of relevant molecular weight bands to PtIP-50 polypeptidesand/or PtIP-65 polypeptides) with sequence information of PtIP-50polypeptides of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO:46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ IDNO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO:65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ IDNO: 70 and/or PtIP-65 polypeptides (e.g., SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ IDNO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO:42)) and its homologs. Any match in peptide sequences indicates thepotential of having the homologous proteins in the samples. Additionaltechniques (protein purification and molecular biology) can be used toisolate the protein and identify the sequences of the homologs.

In hybridization methods, all or part of the pesticidal nucleic acidsequence can be used to screen cDNA or genomic libraries. Methods forconstruction of such cDNA and genomic libraries are generally known inthe art and are disclosed in Sambrook and Russell, (2001), supra. Theso-called hybridization probes may be genomic DNA fragments, cDNAfragments, RNA fragments or other oligonucleotides and may be labeledwith a detectable group such as 32P or any other detectable marker, suchas other radioisotopes, a fluorescent compound, an enzyme or an enzymeco-factor. Probes for hybridization can be made by labeling syntheticoligonucleotides based on the known PtIP-50 polypeptide or PtIP-65polypeptide-encoding nucleic acid sequence disclosed herein. Degenerateprimers designed on the basis of conserved nucleotides or amino acidresidues in the nucleic acid sequence or encoded amino acid sequence canadditionally be used. The probe typically comprises a region of nucleicacid sequence that hybridizes under stringent conditions to at leastabout 12, at least about 25, at least about 50, 75, 100, 125, 150, 175or 200 consecutive nucleotides of nucleic acid sequence encoding aPtIP-50 polypeptide or PtIP-65 polypeptide of the disclosure or afragment or variant thereof. Methods for the preparation of probes forhybridization are generally known in the art and are disclosed inSambrook and Russell, (2001), supra, herein incorporated by reference.

For example, an entire nucleic acid sequence, encoding a PtIP-50polypeptide and/or PtIP-65 polypeptide, disclosed herein or one or moreportions thereof may be used as a probe capable of specificallyhybridizing to corresponding nucleic acid sequences encoding PtIP-50polypeptide or PtIP-65 polypeptide-like sequences and messenger RNAs. Toachieve specific hybridization under a variety of conditions, suchprobes include sequences that are unique and are preferably at leastabout 10 nucleotides in length or at least about 20 nucleotides inlength. Such probes may be used to amplify corresponding pesticidalsequences from a chosen organism by PCR. This technique may be used toisolate additional coding sequences from a desired organism or as adiagnostic assay to determine the presence of coding sequences in anorganism. Hybridization techniques include hybridization screening ofplated DNA libraries (either plaques or colonies; see, for example,Sambrook, et al., (1989) Molecular Cloning: A Laboratory Manual (2d ed.,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).

Hybridization of such sequences may be carried out under stringentconditions. “Stringent conditions” or “stringent hybridizationconditions” is used herein to refer to conditions under which a probewill hybridize to its target sequence to a detectably greater degreethan to other sequences (e.g., at least 2-fold over background).Stringent conditions are sequence-dependent and will be different indifferent circumstances. By controlling the stringency of thehybridization and/or washing conditions, target sequences that are 100%complementary to the probe can be identified (homologous probing).Alternatively, stringency conditions can be adjusted to allow somemismatching in sequences so that lower degrees of similarity aredetected (heterologous probing). Generally, a probe is less than about1000 nucleotides in length, preferably less than 500 nucleotides inlength.

Typically, stringent conditions will be those in which the saltconcentration is less than about 1.5 M Na ion, typically about 0.01 to1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and thetemperature is at least about 30° C. for short probes (e.g., 10 to 50nucleotides) and at least about 60° C. for long probes (e.g., greaterthan 50 nucleotides). Stringent conditions may also be achieved with theaddition of destabilizing agents such as formamide. Exemplary lowstringency conditions include hybridization with a buffer solution of 30to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulphate) at 37° C.,and a wash in 1× to 2×SSC (20×SSC=3.0 M NaCl/0.3 M trisodium citrate) at50 to 55° C. Exemplary moderate stringency conditions includehybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., anda wash in 0.5× to 1×SSC at 55 to 60° C. Exemplary high stringencyconditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at37° C., and a wash in 0.1×SSC at 60 to 65° C. Optionally, wash buffersmay comprise about 0.1% to about 1% SDS. Duration of hybridization isgenerally less than about 24 hours, usually about 4 to about 12 hours.

Specificity is typically the function of post-hybridization washes, thecritical factors being the ionic strength and temperature of the finalwash solution. For DNA-DNA hybrids, the Tm can be approximated from theequation of Meinkoth and Wahl, (1984) Anal. Biochem. 138:267-284:Tm=81.5° C.+16.6 (log M)+0.41 (% GC)−0.61 (% form)−500/L; where M is themolarity of monovalent cations, % GC is the percentage of guanosine andcytosine nucleotides in the DNA, % form is the percentage of formamidein the hybridization solution, and L is the length of the hybrid in basepairs. The Tm is the temperature (under defined ionic strength and pH)at which 50% of a complementary target sequence hybridizes to aperfectly matched probe. Tm is reduced by about 1° C. for each 1% ofmismatching; thus, Tm, hybridization, and/or wash conditions can beadjusted to hybridize to sequences of the desired identity. For example,if sequences with ≥90% identity are sought, the Tm can be decreased 10°C. Generally, stringent conditions are selected to be about 5° C. lowerthan the thermal melting point (Tm) for the specific sequence and itscomplement at a defined ionic strength and pH. However, severelystringent conditions can utilize a hybridization and/or wash at 1, 2, 3or 4° C. lower than the thermal melting point (Tm); moderately stringentconditions can utilize a hybridization and/or wash at 6, 7, 8, 9 or 10°C. lower than the thermal melting point (Tm); low stringency conditionscan utilize a hybridization and/or wash at 11, 12, 13, 14, 15 or 20° C.lower than the thermal melting point (Tm). Using the equation,hybridization and wash compositions, and desired Tm, those of ordinaryskill will understand that variations in the stringency of hybridizationand/or wash solutions are inherently described. If the desired degree ofmismatching results in a Tm of less than 45° C. (aqueous solution) or32° C. (formamide solution), it is preferred to increase the SSCconcentration so that a higher temperature can be used. An extensiveguide to the hybridization of nucleic acids is found in Tijssen, (1993)Laboratory Techniques in Biochemistry and MolecularBiology-Hybridization with Nucleic Acid Probes, Part I, Chapter 2(Elsevier, N.Y.); and Ausubel, et al., eds. (1995) Current Protocols inMolecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience,New York). See, Sambrook, et al., (1989) Molecular Cloning: A LaboratoryManual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y.).

Proteins and Variants and Fragments Thereof

PtIP-50 polypeptides and PtIP-65 polypeptides are also encompassed bythe disclosure. “Pteridophyta Insecticidal Protein-50” “PtIP-50polypeptide”, and “PtIP-50 protein” as used herein interchangeablyrefers to a polypeptide having insecticidal activity including but notlimited to insecticidal activity against one or more insect pests of theLepidoptera and/or Coleoptera orders, and is sufficiently homologous tothe protein of SEQ ID NO: 71. A variety of PtIP-50 polypeptides arecontemplated. Sources of PtIP-50 polypeptides or related proteins arefern species selected from but not limited to Asplenium australasicum,Asplenium nidus, Asplenium×kenzoi Sa. Kurata, Polypodium musifolium,Polypodium punctatum ‘Serratum’, Adiantum pedatum L., Platyceriumbifurcatum, Nephrolepis falcata, Colysis wrightii ‘Monstifera’, Colysiswrightii, Bolbitis cladorrhizans, and Blechnum brasiliense ‘Crispum’.“Pteridophyta Insecticidal Protein-65” “PtIP-65 polypeptide”, and“PtIP-65 protein” as used herein interchangeably refers to a polypeptidehaving insecticidal activity including but not limited to insecticidalactivity against one or more insect pests of the Lepidoptera and/orColeoptera orders, and is sufficiently homologous to the protein of SEQID NO: 22. A variety of PtIP-65 polypeptides are contemplated. Sourcesof PtIP-65 polypeptides or related proteins are fern species selectedfrom but not limited to: Asplenium australasicum, Asplenium×kenzoi Sa.Kurata, Polypodium musifolium, Polypodium punctatum ‘Serratum’, Adiantumpedatum L., Platycerium bifurcatum, Nephrolepis falcata, Ophioglossumpendulum, Colysis wrightii ‘Monstifera’, Colysis wrightii, Selaginellakraussiana ‘Variegata’, Selaginella victoriae, Lycopodium phlehmaria,and Huperzia salvinioides.

“Sufficiently homologous” is used herein to refer to an amino acidsequence that has at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or greater sequence homology compared to a reference sequenceusing one of the alignment programs described herein using standardparameters. In some embodiments the sequence homology is against thefull length sequence of a PtIP-50 polypeptide. In some embodiments thePtIP-50 polypeptide has at least about 40%, 45%, 50%, 51%, 52%, 53%,54%, 55%, 56%, 57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%,68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or greater sequence identity compared to SEQ ID NO:71. In some embodiments the sequence homology is against the full lengthsequence a PtIP-65 polypeptide. In some embodiments the PtIP-65polypeptide has at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or greater sequence identity compared to SEQ ID NO: 22. One ofskill in the art will recognize that these values can be appropriatelyadjusted to determine corresponding homology of proteins taking intoaccount amino acid similarity and the like. In some embodiments thesequence identity is calculated using ClustalW algorithm in the ALIGNX®module of the Vector NTI® Program Suite (Invitrogen Corporation,Carlsbad, Calif.) with all default parameters. In some embodiments thesequence identity is across the entire length of polypeptide calculatedusing ClustalW algorithm in the ALIGNX® module of the Vector NTI®Program Suite (Invitrogen Corporation, Carlsbad, Calif.) with alldefault parameters.

As used herein, the terms “protein,” “peptide molecule,” or“polypeptide” includes any molecule that comprises five or more aminoacids. It is well known in the art that protein, peptide or polypeptidemolecules may undergo modification, including post-translationalmodifications, such as, but not limited to, disulfide bond formation,glycosylation, phosphorylation or oligomerization. Thus, as used herein,the terms “protein,” “peptide molecule” or “polypeptide” includes anyprotein that is modified by any biological or non-biological process.The terms “amino acid” and “amino acids” refer to all naturallyoccurring L-amino acids.

A “recombinant protein” is used herein to refer to a protein that is nolonger in its natural environment, for example in vitro or in arecombinant bacterial or plant host cell. A PtIP-50 polypeptide orPtIP-65 polypeptide that is substantially free of cellular materialincludes preparations of protein having less than about 30%, 20%, 10% or5% (by dry weight) of non-pesticidal protein (also referred to herein asa “contaminating protein”).

“Fragments” or “biologically active portions” include polypeptidefragments comprising amino acid sequences sufficiently identical to aPtIP-50 polypeptide or PtIP-65 polypeptide and that exhibit insecticidalactivity. “Fragments” or “biologically active portions” of PtIP-50polypeptides includes fragments comprising amino acid sequencessufficiently identical to the amino acid sequence set forth in SEQ IDNO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80,SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO:85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ IDNO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98, wherein thePtIP-50 polypeptide, in combination with a PtIP-65 polypeptide, hasinsecticidal activity. “Fragments” or “biologically active portions” ofPtIP-65 polypeptides includes fragments comprising amino acid sequencessufficiently identical to the amino acid sequence set forth in SEQ IDNO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31,SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO:36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ IDNO: 41 or SEQ ID NO: 42, wherein the PtIP-65 polypeptide, in combinationwith a PtIP-50 polypeptide, has insecticidal activity. Such biologicallyactive portions can be prepared by recombinant techniques and evaluatedfor insecticidal activity. In some embodiments, the PtIP-50 polypeptidefragment is an N-terminal and/or a C-terminal truncation of at leastabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more amino acids from theN-terminus and/or C-terminus relative to SEQ ID NO: 71, SEQ ID NO: 72,SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO:77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ IDNO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO:96, SEQ ID NO: 97 or SEQ ID NO: 98 e.g., by proteolysis, by insertion ofa start codon, by deletion of the codons encoding the deleted aminoacids and concomitant insertion of a start codon, and/or insertion of astop codon.

In some embodiments, the PtP-50 polypeptide fragments encompassed hereinresult from the removal of the N-terminal 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 or more amino acids relative to SEQ ID NO: 71, SEQ ID NO: 72, SEQ IDNO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82,SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO:87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ IDNO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQID NO: 97 or SEQ ID NO: 98 or variants thereof, e.g., by proteolysis orby insertion of a start codon, by deletion of the codons encoding thedeleted amino acids and concomitant insertion of a start codon.

“Variants” as used herein refers to proteins or polypeptides having anamino acid sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% identical to the parental amino acidsequence.

PtIP-50 Polypeptides

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 71, SEQ ID NO: 72,SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO:77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ IDNO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO:96, SEQ ID NO: 97 or SEQ ID NO: 98, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identityacross the entire length of the amino acid sequence of SEQ ID NO: 71,SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ IDNO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90,SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO:95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 98.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 71, SEQ IDNO: 77, SEQ ID NO: 78, SEQ ID NO: 84 or SEQ ID NO: 96, wherein thePtIP-50 polypeptide, in combination with a PtIP-65 polypeptide, hasinsecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 71, SEQ ID NO: 77, SEQ ID NO: 78,SEQ ID NO: 84 or SEQ ID NO: 96, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 72, SEQ IDNO: 73, SEQ ID NO: 74, SEQ ID NO: 79, SEQ ID NO: 85, SEQ ID NO: 89, SEQID NO: 93 or SEQ ID NO: 94, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 79, SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 93 or SEQ ID NO:94, wherein the PtIP-50 polypeptide, in combination with a PtIP-65polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 75, SEQ IDNO: 81, SEQ ID NO: 82, SEQ ID NO: 87 or SEQ ID NO: 98, wherein thePtIP-50 polypeptide, in combination with a PtIP-65 polypeptide, hasinsecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 75, SEQ ID NO: 81, SEQ ID NO: 82,SEQ ID NO: 87 or SEQ ID NO: 98, wherein the PtIP-50 polypeptide, incombination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 76, SEQ IDNO: 80, SEQ ID NO: 88 or SEQ ID NO: 92, wherein the PtIP-50 polypeptide,in combination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 76, SEQ ID NO: 80, SEQ ID NO: 88or SEQ ID NO: 92, wherein the PtIP-50 polypeptide, in combination with aPtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 83, SEQ IDNO: 86, SEQ ID NO: 90 or SEQ ID NO: 91, wherein the PtIP-50 polypeptide,in combination with a PtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 83, SEQ ID NO: 86, SEQ ID NO: 90or SEQ ID NO: 91, wherein the PtIP-50 polypeptide, in combination with aPtIP-65 polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 97, whereinthe PtIP-50 polypeptide, in combination with a PtIP-65 polypeptide, hasinsecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity tothe amino acid sequence of SEQ ID NO: 97, wherein the PtIP-50polypeptide, in combination with a PtIP-65 polypeptide, has insecticidalactivity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having at least about 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to any one of the amino acid sequences of SEQ ID NO:224-326, wherein the PtIP-50 polypeptide, in combination with a PtIP-65polypeptide, has insecticidal activity.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO:98, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70 or moreamino acid substitutions compared to the native amino acid at thecorresponding position of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73,SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO:78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ IDNO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92,SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO:97 or SEQ ID NO: 98.

In some embodiments the sequence identity is across the entire length ofthe polypeptide calculated using ClustalW algorithm in the ALIGNX®module of the Vector NTI® Program Suite (Invitrogen Corporation,Carlsbad, Calif.) with all default parameters.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59 or 60 amino acid substitutions, in anycombination, compared to the native amino acid at the correspondingposition of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO:98.

In some embodiments a PtIP-50 polypeptide comprises an amino acidsequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 or 29 amino acidsubstitutions, in any combination, compared to the native amino acid atthe corresponding position of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO:73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ IDNO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO:92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ IDNO: 97 or SEQ ID NO: 98.

In some embodiments the PtIP-50 polypeptide comprises an amino acidsequence of SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO:98.

In some embodiments the nucleic acid molecule encoding the PtIP-50polypeptide is derived from a fern species in the Division Pteridophyta.In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Class Psilotopsida. In some embodiments the PtIP-50polypeptide is derived from a fern species in the Class Psilotopsida,Order Psilotales. In some embodiments the PtIP-50 polypeptide is derivedfrom a fern species in the Class Psilotopsida, Order Ophioglossales. Insome embodiments the PtIP-50 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales, Family Psilotaceae. Insome embodiments the PtIP-50 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales Family Ophioglossaceae.In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Genus Ophioglossum L., Bottychium, Botrypus,Helminthostachys, Ophioderma, Cheiroglossa, Sceptridium or Mankyua. Insome embodiments the PtIP-50 polypeptide is derived from a fern speciesin the Genus Ophioglossum L. is selected from but not limited toOphioglossum califomicum, Ophioglossum coriaceum, Ophioglossum costatum,Ophioglossum crotalophoroides, Ophioglossum engelmannii, Ophioglossumfalcaturn, Ophioglossum gomezianurn, Ophioglossum gramineum,Ophioglossum kawamurae, Ophioglossum lusitanicum, Ophioglossumnamegatae, Ophioglossum nudicaule, Ophioglossum palmatum, Ophioglossumparvum, Ophioglossum pedunculosum, Ophioglossum pendulum, Ophioglossumpetiolatum, Ophioglossum pusillum, Ophioglossum reticulatum,Ophioglossum richardsiae, Ophioglossum thermale, and Ophioglossumvulgatum.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Class Polypodiopsida/Pteridopsida. In some embodiments the PtIP-50polypeptide is derived from a fern species in the Order Osmundales(royal ferns); Family Osmundaceae. In some embodiments the PtIP-50polypeptide is derived from a fern species in the Order Hymenophyllales(filmy ferns and bristle ferns); Family Hymenophyllaceae. In someembodiments the PtIP-50 polypeptide is derived from a fern species inthe Order Gleicheniales; Family Gleicheniaceae, Family Dipteridaceael orFamily Matoniaceae. In some embodiments the PtIP-50 polypeptide isderived from a fern species in the Order Schizaeales; FamilyLygodiaceae, Family Anemiaceae or Family Schizaeaceae. In someembodiments the PtIP-50 polypeptide is derived from a fern species inthe Order Salviniales; Family Marsileaceae or Family Salviniaceae. Insome embodiments the PtIP-50 polypeptide is derived from a fern speciesin the Order Cyatheales; Family Thyrsopteridaceae, Family Loxsomataceae,Family Culcitaceae, Family Plagiogyriaceae, Family Cibotiaceae, FamilyCyatheaceae, Family Dicksoniaceae or Family Metaxyaceae.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales; Family Lindsaeaceae, FamilySaccolomataceae, Family Cystodiaceae, Family Dennstaedtiaceae, FamilyPteridaceae, Family Aspleniaceae, Family Thelypteridaceae, FamilyWoodsiaceae, Family Onocleaceae, Family Blechnaceae, FamilyDryopteridaceae, Family Lomariopsidaceae, Family Tectariaceae, FamilyOleandraceae, Family Davalliaceae or Family Polypodiaceae.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Pteridaceae, Genus Adiantaceaeselected from but not limited to Adiantum aethiopicum, Adiantumaleuticum, Adiantum bonatianum, Adiantum cajennense, Adiantumcapillus-junonis, Adiantum capillus-veneris, Adiantum caudatum, Adiantumchienfi, Adiantum chilense, Adiantum cuneatum, Adiantum cunninghamfi,Adiantum davidii, Adiantum diaphanum, Adiantum edentulum, Adiantumedgeworthfi, Adiantum excisum, Adiantum fengianum, Adiantum fimbriatum,Adiantum flabellulaturn, Adiantum formosanum, Adiantum formosum,Adiantum fulvum, Adiantum gravesfi, Adiantum hispidulum, Adiantuminduratum, Adiantum jordanfi, Adiantum juxtapositum, Adiantumlatifolium, Adiantum leveillei, Adiantum lianxianense, Adiantummalesianum, Adiantum mariesfi, Adiantum monochlamys, Adiantummyriosorum, Adiantum obliquum, Adiantum ogasawarense, Adiantum pedatum,Adiantum pentadactylon, Adiantum peruvianum, Adiantum philippense,Adiantum princeps, Adiantum pubescens, Adiantum raddianum, Adiantumreniforme, Adiantum roborowskii, Adiantum serratodentatum, Adiantumsinicum, Adiantum soboliferum, Adiantum subcordatum, Adiantum tenerum,Adiantum terminatum, Adiantum tetraphyllum, Adiantum venustum, Adiantumviridescens, and Adiantum viridimontanurn.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL. In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL selected from but not limited to Asplenium abbreviatum, Aspleniumabrotanoides, Asplenium abscissum var. subaequilaterale, Aspleniumabscissum, Asplenium achilleifolium, Asplenium acuminatum, Aspleniumadiantifrons, Asplenium adiantoides, Asplenium adiantoides var.squamulosum, Asplenium adiantum-nigrum L., Asplenium adiantum-nigrumvar. adiantum-nigrum, Asplenium adiantum-nigrum var. yuanurn, Aspleniumadnatum, Asplenium aethiopicum, Asplenium affine, Asplenium affine var.affine, Asplenium affine var. gilpinae, Asplenium affine var. mettenii,Asplenium affine var. pecten, Asplenium africanum, Asplenium afzelii,Asplenium aitchisonii, Asplenium alatulum, Asplenium alatum, Aspleniumalfredii, Asplenium altajense, Asplenium amabile, Aspleniumambohitantelense, Asplenium anceps var. proliferum, Asplenium andapense,Asplenium andersonii, Asplenium angustatum, Asplenium angustum,Asplenium anisophyllum, Asplenium annetii, Asplenium antiquum, Aspleniumantrophyoides, Asplenium apertura, Asplenium apogamum, Aspleniumaquaticum, Asplenium arboreum, Asplenium arcanum, Asplenium arcuatum,Asplenium argentinurn, Asplenium argutum, Asplenium aspidiiforme,Asplenium aspidioides, Asplenium asterolepis, Asplenium auriculariumvar. acutidens, Asplenium auricularium var. subintegerrimum, Aspleniumauriculatum, Asplenium auriculaturn var. aequilaterale, Aspleniumauritum fo. diversifolium, Asplenium auritum fo. diversifolium,Asplenium auritum fo. nana, Asplenium auritum, Asplenium auritum var.auriculaturn, Asplenium auritum var. auritum, Asplenium auritum var.bipinnatifidum, Asplenium auritum var. bipinnatisectum, Aspleniumauritum var. davallioides, Asplenium auritum var. macilentum, Aspleniumauritum var. rigidum, Asplenium auritum var. subsimplex, Aspleniumaustrochinense, Asplenium ayopayense, Asplenium badinii, Aspleniumbalense, Asplenium ballivianii, Asplenium bangii, Asplenium bangii,Asplenium barbaense, Asplenium barclayanum, Asplenium barkamense,Asplenium barteri, Asplenium basiscopicum, Asplenium bicrenaturn,Asplenium bifrons, Asplenium bipartitum, Asplenium blastophorum,Asplenium blepharodes, Asplenium blepharophorum, Asplenium boiteaui,Asplenium bolivianum, Asplenium boltonii, Asplenium borealichinense,Asplenium bradei, Asplenium bradeorum, Asplenium bradleyi, Aspleniumbrausei, Asplenium breedlovei, Asplenium buettneri, Asplenium buettnerivar. hildebrandtii, Asplenium bulbiferum, Asplenium bullaturn var.bullaturn, Asplenium bullatum var. shikokianum, Asplenium bullaturn,Asplenium cancellatum, Asplenium capillipes, Asplenium cardiophyllum(Hance), Asplenium caripense, Asplenium carvalhoanum, Aspleniumcastaneoviride, Asplenium castaneum, Asplenium caudatum, Aspleniumceltidifolium (Kunze), Asplenium ceratolepis, Asplenium changputungense,Asplenium chaseanum, Asplenium cheilosorum, Asplenium chengkouense,Asplenium chihuahuense, Asplenium chimantae, Asplenium chimborazense,Asplenium chingianum, Asplenium chlorophyllum, Asplenium chondrophyllum,Asplenium cicutarium, Asplenium cicutarium var. paleaceum, Aspleniumcirrhaturn, Asplenium cladolepton, Asplenium claussenii, Aspleniumcoenobiale, Asplenium commutatum, Asplenium congestum, Aspleniumconquisitum, Asplenium consimile, Asplenium contiguum, Aspleniumcontiguum var. hirtulum, Asplenium corderoi, Asplenium cordovense,Asplenium coriaceum, Asplenium corfifolium, Asplenium correardii,Asplenium costale, Asplenium costale var. robustum, Asplenium cowanii,Asplenium crenulatoserrulatum, Asplenium crenulatum, Aspleniumcrinicaule, Asplenium crinulosum, Asplenium cristatum, Aspleniumctyptolepis Fernald, Asplenium cultrifolium L., Asplenium cuneatiforme,Asplenium cuneatum, Asplenium curvatura, Asplenium cuspidatum, Aspleniumcuspidatum var cuspidatum, Asplenium cuspidatum var. foeniculaceum,Asplenium cuspidatum var. triculum, Asplenium cuspidatum var.tripinnaturn, Asplenium dalhousiae, Asplenium dareoides, Aspleniumdavallioides, Asplenium davisii, Asplenium debile, Asplenium debile,Asplenium decussatum, Asplenium delavayi, Asplenium delicatulum,Asplenium delicatulum var. cocosensis, Asplenium delitescens, Aspleniumdelitescens×laetum, Asplenium densum, Asplenium dentatum L., Aspleniumdentatum L., Asplenium depauperatum, Asplenium deqenense, Aspleniumdianae, Asplenium difforme, Asplenium dilatatum, Asplenium dimidiatum,Asplenium dimidiaturn var. boliviense, Asplenium diplazisorum, Aspleniumdissectum, Asplenium distans, Asplenium divaricatum, Aspleniumdivergens, Asplenium divisissimurn, Asplenium doederleinii, Aspleniumdonnell-smithii, Asplenium dregeanum, Asplenium dulongjiangense,Asplenium duplicatoserratum, Asplenium eatonii, Asplenium ebeneum,Asplenium ebenoides, Asplenium ecuadorense, Asplenium eggersii,Asplenium emarginaturn, Asplenium enaturn, Asplenium ensiforme fo.bicuspe, Asplenium ensiforme fo. ensiforme, Asplenium ensiforme fo.stenophyllum, Asplenium ensiforme, Asplenium erectum var. erectum,Asplenium erectum var. gracile, Asplenium erectum var. usambarense,Asplenium erectum var. zeyheri, &, Asplenium erosum L., Aspleniumescaleroense, Asplenium esculenturn, Asplenium eutecnurn, Aspleniumexcelsum, Asplenium excisum, Asplenium exiguum, Asplenium extensum,Asplenium falcatum, Asplenium falcinellum, Asplenium faurei, Aspleniumfeei, Asplenium fengyangshanense, Asplenium ferulaceum, Aspleniumfibrillosum, Asplenium filix-femina, Asplenium finckii, Aspleniumfinlaysonianurn, Asplenium flabellulatum, Asplenium flabellulatum varflabellulatum, Asplenium flabellulatum var. partitum, Aspleniumflaccidum, Asplenium flavescens, Asplenium flavidum, Aspleniumflexuosum, Asplenium fluminense, Asplenium foeniculaceum, Aspleniumformosanum, Asplenium formosum var. carolinurn, Asplenium formosum var.incultum, Asplenium formosum, Asplenium fournieri, Asplenium fragile,Asplenium fragile var. lomense, Asplenium fragrans, Asplenium fragransvar. foeniculaceum, Asplenium franconis var. gracile, Aspleniumfraxinifolium, Asplenium friesiorum, Asplenium friesiorum var.nesophilum, Asplenium fugax, Asplenium fujianense, Asplenium furcatum,Asplenium furfuraceum, Asplenium fuscipes, Asplenium fuscopubescens,Asplenium galeottii, Asplenium gautieri, Asplenium gemmiferum, Aspleniumgentryi, Asplenium geppii, Asplenium ghiesbreghtii, Asplenium gilliesii,Asplenium gilpinae, Asplenium glanduliserratum, Asplenium glenniei,Asplenium goldmannii, Asplenium gomezianum, Asplenium grande, Aspleniumgrandifolium, Asplenium grandifrons, Asplenium gregoriae, Aspleniumgriffithianum, Asplenium gulingense, Asplenium hainanense, Aspleniumhallbergii, Asplenium hallei, Asplenium hallii, Asplenium hangzhouense,Asplenium haplophyllum, Asplenium harpeodes, Asplenium harpeodes var.glaucovirens, Asplenium harpeodes var. incisura, Asplenium harrisiiJenman, Asplenium harrisonii, Asplenium hastatum, Asplenium hebeiense,Asplenium hemionitideum, Asplenium hemitomurn, Asplenium henryi,Asplenium herpetopteris, Asplenium herpetopteris var herpetopteris,Asplenium herpetopteris var. acutipinnata, Asplenium herpetopteris var.masoulae, Asplenium herpetopteris var. villosum, Asplenium hesperium,Asplenium heterochroum, Asplenium hians, Asplenium hians var.pallescens, Asplenium hoffmannii, Asplenium holophlebium, Aspleniumhondoense, Asplenium horridum, Asplenium hostmannii, Aspleniumhumistratum, Asplenium hypomelas, Asplenium inaequilaterale, Aspleniumincisura, Asplenium incurvaturn, Asplenium indicum, Asplenium indicumvar. indicum, Asplenium indicum var. yoshingagae, Asplenium induratum,Asplenium indusiatum, Asplenium inexpectatum, Asplenium insigne,Asplenium insiticium, Asplenium insolitum, Asplenium integerrimurn,Asplenium interjectum, Asplenium jamesonii, Asplenium jaundeense,Asplenium juglandifolium, Asplenium kangdingense, Asplenium kansuense,Asplenium kassneri, Asplenium kaulfussii, Asplenium kellermanii,Asplenium kentuckiense, Asplenium khullarii, Asplenium kiangsuense,Asplenium kunzeanurn, Asplenium lacerum, Asplenium laciniatum, Aspleniumlaciniatum var. acutipinna, Asplenium laciniatum var. laciniatum,Asplenium laetum fo. minor, Asplenium laetum, Asplenium laetum var.incisoserratum, Asplenium lamprocaulon, Asplenium laserpitiifolium var.morrisonense, Asplenium lastii, Asplenium latedens, Aspleniumlatifolium, Asplenium laui, Asplenium laurentii, Asplenium leandrianurn,Asplenium lechleri, Asplenium leiboense, Asplenium lepidorachis,Asplenium leptochlamys, Asplenium leptophyllum, Asplenium levyi,Asplenium lindbergii, Asplenium lindeni, Asplenium lineatum, Aspleniumlividum, Asplenium lobatum, Asplenium lobulaturn, Asplenium lokohoense,Asplenium longicauda, Asplenium longicaudaturn, Asplenium longifolium,Asplenium longisorum, Asplenium longjinense, Asplenium lorentzii,Asplenium loriceum, Asplenium loxogrammoides, Asplenium lugubre,Asplenium lunulatum, Asplenium lunulatum var. pteropus, Aspleniumlushanense, Asplenium lydgatei, Asplenium macilentum, Asplenium macraei,Asplenium macrodictyon, Asplenium macrophlebium, Asplenium macrophyllum,Asplenium macropterum, Asplenium macrosorum, Asplenium macrotis,Asplenium macrurum, Asplenium mainlingense, Asplenium mangindranense,Asplenium mannii, Asplenium marginatum L., Asplenium marojejyense,Asplenium martianum, Asplenium matsumurae, Asplenium mauritiensisLorence, Asplenium maximum, Asplenium, ii, Asplenium megalura, Aspleniummegaphyllum, Asplenium meiotomurn, Asplenium melanopus, Aspleniummembranifolium, Asplenium meniscioides, Asplenium mesosorum, Aspleniummexicanurn, Asplenium micropaleatum, Asplenium microtum, Aspleniummildbraedii, Asplenium mildei, Asplenium minimum, Asplenium minutum,Asplenium miradorense, Asplenium miyunense, Asplenium moccenianum,Asplenium mocquetysii, Asplenium modestum, Asplenium monanthemum var.menziesii, Asplenium monanthes L., Asplenium monanthes var monanthes,Asplenium monanthes var. castaneum, Asplenium monanthes var. wagneri,Asplenium monanthes var. yungense, Asplenium monodon, Aspleniummontanum, Asplenium mosetenense, Asplenium moupinense, Aspleniummucronatum, Asplenium munchii, Asplenium muticum, Asplenium myapteron,Asplenium myriophyllu, Asplenium nakanoanum, Asplenium nanchuanense,Asplenium nemorale, Asplenium neolaserpitiifolium, Aspleniumneomutijugum, Asplenium neovarians, Asplenium nesii, Aspleniumnesioticum, Asplenium nidus L., Asplenium nigricans, Aspleniumniponicum, Asplenium normale, Asplenium normale var. angustum, Aspleniumobesum, Asplenium oblongatum, Asplenium oblongifolium, Aspleniumobovatum, Asplenium obscurum, Asplenium obscurum var. angustum,Asplenium obtusatum var. obtusatum, Asplenium obtusatum var. sphenoides,Asplenium obtusifolium L., Asplenium obtusissimum, Asplenium obversum,Asplenium ochraceum, Asplenium oellgaardii, Asplenium ofeliae, Aspleniumoldhami, Asplenium oligosorum, Asplenium olivaceum, Asplenium onopterisL., Asplenium onustum, Asplenium ortegae, Asplenium otites, Aspleniumpalaciosii, Asplenium palmeri, Asplenium partitum, Asplenium parvisorum,Asplenium parviusculum, Asplenium parvulum, Asplenium patens, Aspleniumpaucifolium, Asplenium paucijugum, Asplenium paucivenosum, Aspleniumpearcei, Asplenium pekinense, Asplenium pellucidum, Asplenium pendulum,Asplenium petiolulatum, Asplenium phyllitidis, Aspleniumpimpinellifolium, Asplenium pinnatifidum, Asplenium pinnatum, Aspleniumplatyneuron, Asplenium platyneuron var. bacculum-rubrum, Aspleniumplatyneuron var. incisum, Asplenium platyphyllum, Asplenium plumbeum,Asplenium poloense, Asplenium polymeris, Asplenium polymorphum,Asplenium polyodon, Asplenium polyodon var. knudsenii, Aspleniumpolyodon var. nitidulum, Asplenium polyodon var. sectum, Aspleniumpolyodon var. subcaudatum, Asplenium polyphyllum, Asplenium poolii,Asplenium poolii fo. simplex, Asplenium poolii var. linearipinnatum,Asplenium potosinum, Asplenium potosinum var. incisum, Aspleniumpraegracile, Asplenium praemorsum, Asplenium preussii, Aspleniumpringleanum, Asplenium pringlei, Asplenium prionitis, Aspleniumprocerum, Asplenium progrediens, Asplenium projectum, Aspleniumprolongatum, Asplenium propinquum, Asplenium protensum, Aspleniumpseudoangustum, Asplenium pseudoerectum, Asplenium pseudofontanum,Asplenium pseudolaserpitiifolium, Asplenium pseudonormale, Aspleniumpseudo pellucidum, Asplenium pseudopraemorsum, Asplenium pseudovarians,Asplenium pseudowilfordii, Asplenium pseudowrightii, Aspleniumpsilacrum, Asplenium pteropus, Asplenium pubirhizoma, Aspleniumpulchellum, Asplenium pulchellum var. subhorizontale, Aspleniumpulcherrimum, Asplenium pulicosum, Asplenium pulicosum var. maius,Asplenium pululahuae, Asplenium pumilum, Asplenium pumilum var.hymenophylloides, Asplenium pumilum var. laciniatum, Aspleniumpurdieanum, Asplenium purpurascens, Asplenium pyramidatum, Aspleniumqiujiangense, Asplenium quercicola, Asplenium quitense, Aspleniumraddianum, Asplenium radiatum, Asplenium radicans L., Aspleniumradicans, Asplenium radicans var. costaricense, Asplenium radicans var.partitum, Asplenium radicans var. radicans, Asplenium radicans var.uniseriale, Asplenium recumbens, Asplenium reflexum, Asplenium regularevar. latior, Asplenium repandulum, Asplenium repens, Asplenium repente,Asplenium resiliens, Asplenium retusulum, Asplenium rhipidoneuron,Asplenium rhizophorum L., Asplenium rhizophyllum, Asplenium rhizophyllumL., Asplenium rhizophyllum var. proliferum, Asplenium rhomboideum,Asplenium rigidum, Asplenium riparium, Asplenium rivale, Aspleniumrockii, Asplenium roemerianum, Asplenium roemerianum var. mindensis,Asplenium rosenstockianum, Asplenium rubinum, Asplenium ruizianum,Asplenium rusbyanum, Asplenium ruta-muraria L., Asplenium ruta-murariavar. cryptolepis, Asplenium rutaceum, Asplenium rutaceum var.disculiferum, Asplenium rutaefolium, Asplenium rutifolium, Aspleniumsalicifolium L., Asplenium salicifolium var. aequilaterale, Aspleniumsalicifolium var. salicifolium, Asplenium sampsoni, Asplenium sanchezii,Asplenium sanderi, Asplenium sandersonii, Asplenium sanguinolentum,Asplenium sarelii, Asplenium sarelii var. magnum, Asplenium sarelii var.sarelii, Asplenium saxicola, Asplenium scalifolium, Aspleniumscandicinum, Asplenium schizophyllum, Asplenium schkuhrii, Aspleniumsciadophilum, Asplenium scolopendrium L., Asplenium scortechinii,Asplenium seileri, Asplenium semipinnatum, Asplenium septentrionale,Asplenium serra, Asplenium serra var. imrayanum, Aspleniumserratissimum, Asplenium serratum L., Asplenium serratum var. caudatum,Asplenium serricula, Asplenium sessilifolium, Asplenium sessilifoliumvar. guatemalense, Asplenium sessilifolium var. minus, Aspleniumsessilifolium var. occidentale, Asplenium sessilipinnum, Aspleniumsetosum, Asplenium shepherdii, Asplenium shepherdii var. bipinnatum,Asplenium shepherdii var. flagelliferum, Asplenium shikokianum,Asplenium simii, Asplenium simonsianum, Asplenium sintenisii, Aspleniumskinneri, Asplenium skinneri, Asplenium sodiroi, Aspleniumsoleirolioides, Asplenium solidum var. stenophyllum, Asplenium solmsii,Asplenium sp.-N.-Halle-2234, Asplenium spathulinum, Aspleniumspectabile, Asplenium speluncae, Asplenium sphaerosporum, Aspleniumsphenotomum, Asplenium spinescens, Asplenium splendens, Aspleniumsprucei, Asplenium squamosum L., Asplenium standleyi, Aspleniumstellatum, Asplenium stenocarpum, Asplenium stoloniferum, Aspleniumstolonipes, Asplenium striatum L., Asplenium stuebelianum, Aspleniumstuhlmannii, Asplenium suave, Asplenium subalatum, Aspleniumsubcrenaturn, Asplenium subdigitatum, Asplenium subdimidiatum, Aspleniumsubintegrum, Asplenium sublaserpitfifolium, Asplenium sublongum,Asplenium subnudum, Asplenium suborbiculare, Asplenium subtenuifolium,Asplenium subtile, Asplenium subtoramanurn, Asplenium subtrapezoideum,Asplenium subvarians, Asplenium sulcatum, Asplenium sylvaticum,Asplenium szechuanense, Asplenium taiwanense, Asplenium tenerrimum,Asplenium tenerum, Asplenium tenuicaule, Asplenium tenuifolium,Asplenium tenuifolium var. minor, Asplenium tenuifolium var.tenuifolium, Asplenium tenuissimum, Asplenium ternatum, Aspleniumtheciferum, Asplenium theciferum var. concinnum, Asplenium thunbergii,Asplenium tianmushanense, Asplenium tianshanense, Asplenium tibeticum,Asplenium tocoraniense, Asplenium toramanurn, Asplenium trapezoideum,Asplenium tricholepis, Asplenium trichomanes L., Asplenium trichomanessubsp. inexpectans, Asplenium trichomanes subsp. quadrivalens, Aspleniumtrichomanes subsp. trichomanes, Asplenium trichomanes var. harovii,Asplenium trichomanes var. herbaceum, Asplenium trichomanes var. repens,Asplenium trichomanes var. viridissimum, Asplenium trichomanes-dentatumL., Asplenium trigonopterum, Asplenium trilobatum, Asplenium trilobum,Asplenium triphyllum, Asplenium triphyllum var. cornpactum, Aspleniumtriphyllum var. gracillimum, Asplenium triphyllum var. herbaceum,Asplenium tripteropus, Asplenium triquetrum, Asplenium truncorum,Asplenium tsaratananense, Asplenium tucumanense, Asplenium tuerckheimii,Asplenium tunquiniense, Asplenium ulbrichtii, Asplenium ultimurn,Asplenium unilaterale, Asplenium unilaterale var. decurrens, Aspleniumunilaterale var. udum, Asplenium unilaterale var. unilaterale, Aspleniumuniseriale, Asplenium uropteron, Asplenium vagans, Aspleniumvareschianum, Asplenium variabile var. paucijugum, Asplenium variabilevar. variabile, Asplenium varians subsp. fimbriatum, Asplenium varians,Asplenium vastum, Asplenium venturae, Asplenium venulosum, Aspleniumverapax, Asplenium vesiculosum, Asplenium vespertinurn, Aspleniumvillosum, Asplenium virens, Asplenium viride, Asplenium viridifrons,Asplenium virillae, Asplenium viviparioides, Asplenium viviparum,Asplenium viviparum var viviparum, Asplenium viviparum var. lineatu,Asplenium volubile, Asplenium vulcanicum, Asplenium wacketii, Aspleniumwagneri, Asplenium wallichianum, Asplenium wameckei, Aspleniumwilfordii, Asplenium williamsii, Asplenium wrightii, Aspleniumwrightioides, Asplenium wuliangshanense, Asplenium xianqianense,Asplenium xinjiangense, Asplenium xinyiense, Asplenium yelagagense,Asplenium yoshinagae, Asplenium yunnanense, Asplenium zamiifolium,Asplenium zanzibaricum, Asplenium biscayneanum, Asplenium curtissii,Asplenium ebenoides, Asplenium herb-wagneri, Asplenium heteroresiliens,Asplenium kenzoi, Asplenium plenum, Asplenium wangii, andAsplenium×clermontiae, Asplenium×gravesii.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.selected from but not limited to Blechnum amabile, Blechnumappendiculatum, Blechnum articulatum, Blechnum australe, Blechnumaustrobrasilianum, Blechnum binervatum, Blechnum blechnoides, Blechnumbrasiliense, Blechnum capense, Blechnum cartilagineum, Blechnumcastaneum, Blechnum chambersii, Blechnum chilense, Blechnum colensoi,Blechnum contiguum, Blechnum cordatum, Blechnum coriaceum, Blechnumdiscolor, Blechnum doodioides, Blechnum durum, Blechnum eburneum,Blechnum ensiforme, Blechnum filiforme, Blechnum fluviatile, Blechnumfragile, Blechnum fraseri, Blechnum fullagari, Blechnum gibbum, Blechnumglandulosum, Blechnum gracile, Blechnum hancockii, Blechnum hastatum,Blechnum howeanum, Blechnum indicum, Blechnum kunthianum, Blechnumlaevigatum, Blechnum loxense, Blechnum magellanicum, Blechnummembranaceum, Blechnum microbasis, Blechnum microphyllum, Blechnummilnei, Blechnum minus, Blechnum mochaenum, Blechnum montanum, Blechnummoorei, Blechnum moritzianum, Blechnum nigrum, Blechnum niponicum,Blechnum norfolkianum, Blechnum novae-zelandiae, Blechnum nudum,Blechnum obtusatum, Blechnum occidentale, Blechnum oceanicum, Blechnumorientate, Blechnum patersonii, Blechnum penna-marina, Blechnumpolypodioides, Blechnum procerum, Blechnum punctulatum, Blechnumsampaioanum, Blechnum schiedeanum, Blechnum schomburgkii, Blechnumserrulatum, Blechnum simillimum, Blechnum spicant, Blechnumstipitellatum, Blechnum tabulare, Blechnum triangularifolium, Blechnumvieillardii, Blechnum vulcanicum, Blechnum wattsii, Blechnum whelanii,and Blechnum wurunuran.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Dryopteridaceae GenusAcrophorus, Genus Acrorumohra, Genus Anapausia, Genus Arachniodes, GenusBolbitis, Genus Ctenitis, Genus Cyclodium, Genus Cyrtogonellum, GenusCyrtomidictyum, Genus Cyrtomium, Genus Diacalpe, Genus Didymochlaena,Genus Dryopsis, Genus Dryopteris, Genus Elaphoglossum, GenusHypodematium, Genus Lastreopsis, Genus Leptorumohra, Genus Leucostegia,Genus Lithostegia, Genus Lomagramma, Genus Maxonia, Genus Megalastrum,Genus Olfersia, Genus Peranema, Genus Phanerophlebia, GenusPhanerophlebiopsis, Genus Polybotrya, Genus Polystichopsis, GenusPolystichum, Genus Rumohra, Genus Sorolepidium, Genus Stigmatopteris orGenus Teratophyllum.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Dryopteridaceae, GenusBolbitis selected from but not limited to Bolbitis acrostichoides,Bolbitis aliena, Bolbitis angustipinna, Bolbitis appendiculata, Bolbitisauriculata, Bolbitis bemoullii, Bolbitis bipinnatifida, Bolbitiscadieri, Bolbitis christensenfi, Bolbitis confertifolia, Bolbitiscostata, Bolbitis crispatula, Bolbitis fluviatilis, Bolbitisgaboonensis, Bolbitis gemmifera, Bolbitis hainanensis, Bolbitis hastata,Bolbitis hekouensis, Bolbitis hemiotis, Bolbitis heteroclita, Bolbitisheudelotii, Bolbitis humblotii, Bolbitis interlineata, Bolbitislatipinna, Bolbitis laxireticulata, Bolbitis lindigii, Bolbitislonchophora, Bolbitis longiflagellata, Bolbitis major, Bolbitis media,Bolbitis nicotianifolia, Bolbitis nodiflora, Bolbitis novoguineensis,Bolbitis oligarchica, Bolbitis palustris, Bolbitis pandurifolia,Bolbitis pergamentacea, Bolbitis portoricensis, Bolbitis presliana,Bolbitis quoyana, Bolbitis rawsonfi, Bolbitis repanda, Bolbitisrhizophylla, Bolbitis riparia, Bolbitis rivularis, Bolbitis sagenioides,Bolbitis salicina, Bolbitis scalpturata, Bolbitis scandens, Bolbitissemicordata, Bolbitis semipinnatifida, Bolbitis serrata, Bolbitisserratifolia, Bolbitis simplex, Bolbitis sinensis, Bolbitissingaporensis, Bolbitis sinuata, Bolbitis subcordata, Bolbitissubcrenata, Bolbitis taylorii, Bolbitis tibetica, Bolbitis tonkinensis,Bolbitis umbrosa, Bolbitis vanuaensis, and Bolbitis virens.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis selected from but not limited to Nephrolepis abrupta,Nephrolepis acuminata, Nephrolepis acutifolia, Nephrolepis arida,Nephrolepis arthropteroides, Nephrolepis biserrata var. auriculata,Nephrolepis brownii, Nephrolepis celebica, Nephrolepis clementis,Nephrolepis cordifolia, Nephrolepis davalliae, Nephrolepis davallioides,Nephrolepis dayakorum, Nephrolepis delicatula, Nephrolepisdicksonioides, Nephrolepis duffii, Nephrolepis exaltata ssp. exaltatassp. Hawaiiensis, Nephrolepis falcata, Nephrolepis falciformis,Nephrolepis glabra, Nephrolepis hirsutula, Nephrolepis humatoides,Nephrolepis iridescens, Nephrolepis kurotawae, Nephrolepis laurifolia,Nephrolepis lauterbachii, Nephrolepis lindsayae, Nephrolepis multifida,Nephrolepis multiflora, Nephrolepis niphoboloides, Nephrolepisobliterate, Nephrolepis paludosa, Nephrolepis pectinata, Nephrolepispendula, Nephrolepis persicifolia, Nephrolepis pickelii, Nephrolepispilosula, Nephrolepis pubescens, Nephrolepis pumicicola, Nephrolepisradicans, Nephrolepis rivularis, Nephrolepis rosenstockii, Nephrolepissaligna, Nephrolepis schlechteri, Nephrolepis serrate, Nephrolepisthomsoni, Nephrolepis undulate var. aureoglandulosa, Nephrolepis×averyi,Nephrolepis×copelandii, and Nephrolepis×medlerae.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusCampyloneurum, Genus Drynaria, Genus Lepisorus, Genus Microgramma, GenusMicrosorum, Genus Neurodium, Genus Niphidium, Genus Pecluma M.G., GenusPhlebodium, Genus Phymatosorus, Genus Platycerium, Genus Pleopeltis,Genus Polypodium L.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L. selected from but not limited to Polypodium absidatum,Polypodium acutifolium, Polypodium adiantiforme, Polypodium aequale,Polypodium affine, Polypodium albidopaleatum, Polypodium alcicorne,Polypodium alfarii, Polypodium alfredii, Polypodium alfredii var.curtii, Polypodium allosuroides, Polypodium alsophilicola, Polypodiumamamianum, Polypodium amoenum, Polypodium amorphum, Polypodiumanetioides, Polypodium anfractuosum, Polypodium anguinum, Polypodiumangustifolium f. remotifolia, Polypodium angustifolium var. amphostenon,Polypodium angustifolium var. heterolepis, Polypodium angustifolium var.monstrosa, Polypodium angustipaleatum, Polypodium angustissimum,Polypodium anisomeron var. pectinatum, Polypodium antioquianum,Polypodium aoristisorum, Polypodium apagolepis, Polypodium apicidens,Polypodium apiculatum, Polypodium apoense, Polypodium appalachianum,Polypodium appressum, Polypodium arenarium, Polypodium argentinurn,Polypodium argutum, Polypodium armaturn, Polypodium aromaticum,Polypodium aspersum, Polypodium assurgens, Polypodium atrum, Polypodiumauriculaturn, Polypodium balaonense, Polypodium balliviani, Polypodiumbamleri, Polypodium bangii, Polypodium bartlettii, Polypodium basale,Polypodium bemoullii, Polypodium biauritum, Polypodium bifrons,Polypodium blepharodes, Polypodium bolivari, Polypodium bolivianurn,Polypodium bolobense, Polypodium bombycinum, Polypodium bombycinum var.insularum, Polypodium bradeorum, Polypodium biyophilum, Polypodiumbtyopodum, Polypodium buchtienii, Polypodium buesii, Polypodiumbulbotrichum, Polypodium caceresii, Polypodium califomicum f.brauscombii, Polypodium califomicum f. parsonsiae, Polypodiumcalifomicum, Polypodium calophlebium, Polypodium calvum, Polypodiumcamptophyllarium var. abbreviatum, Polypodium capitellatum, Polypodiumcarpinterae, Polypodium chachapoyense, Polypodium chartaceum, Polypodiumchimantense, Polypodium chiricanum, Polypodium choquetangense,Polypodium christensenii, Polypodium christii, Polypodium chrysotrichum,Polypodium ciliolepis, Polypodium cinerascens, Polypodium collinsii,Polypodium colysoides, Polypodium confluens, Polypodium conforme,Polypodium confusum, Polypodium congregatifolium, Polypodium connellii,Polypodium consimile var. bourgaeanum, Polypodium consimile var. minor,Polypodium conterminans, Polypodium contiguum, Polypodium cookii,Polypodium coriaceum, Polypodium coronans, Polypodium costaricense,Polypodium costatum, Polypodium crassifolium f. angustissimum,Polypodium crassifolium var. longipes, Polypodium crassulum, Polypodiumcraterisorum, Polypodium cryptum, Polypodium crystalloneuron, Polypodiumcucullatum var. planum, Polypodium cuencanum, Polypodium cumingianum,Polypodium cupreolepis, Polypodium curranii, Polypodium curvans,Polypodium cyathicola, Polypodium cyathisorum, Polypodium cyclocolpon,Polypodium daguense, Polypodium damunense, Polypodium dareiformioides,Polypodium dasypleura, Polypodium decipiens, Polypodium decorum,Polypodium delicatulum, Polypodium deltoideum, Polypodium demeraranum,Polypodium denticulatum, Polypodium diaphanum, Polypodium dilataturn,Polypodium dispersum, Polypodium dissectum, Polypodium dissimulans,Polypodium dolichosorum, Polypodium dolorense, Polypodiumdonnell-smithii, Polypodium drymoglossoides, Polypodium ebeninum,Polypodium eggersii, Polypodium elmeri, Polypodium elongatum, Polypodiumenterosoroides, Polypodium erubescens, Polypodium erythrolepis,Polypodium erythrotrichum, Polypodium eurybasis, Polypodium eurybasisvar. villosum, Polypodium exornans, Polypodium falcoideum, Polypodiumfallacissimum, Polypodium farinosum, Polypodium faucium, Polypodium feetPolypodium ferrugineum, Polypodium feuillei, Polypodium firmulum,Polypodium firmum, Polypodium flaccidum, Polypodium flagellare,Polypodium flexuosum, Polypodium flexuosum var. ekmanii, Polypodiumforbesii, Polypodium formosanum, Polypodium fraxinifolium subsp.articulatum, Polypodium fraxinifolium subsp. luridum, Polypodiumfructuosum, Polypodium fucoides, Polypodium fulvescens, Polypodiumgaleottii, Polypodium glaucum, Polypodium glycyrrhiza, Polypodiumgracillimum, Polypodium gramineum, Polypodium grandifolium, Polypodiumgratum, Polypodium graveolens, Polypodium griseo-nigrum, Polypodiumgriseum, Polypodium guttatum, Polypodium haalilioanum, Polypodiumhammatisorum, Polypodium hancockii, Polypodium haplophlebicum,Polypodium harrisii, Polypodium hastatum var. simplex, Polypodiumhawaiiense, Polypodium heanophyllum, Polypodium helleri, Polypodiumhemionitidium, Polypodium henryi, Polypodium herzogii, Polypodiumhesperium, Polypodium hessii, Polypodium hombersleyi, Polypodiumhostmannii, Polypodium humile, Polypodium hyalinum, Polypodium iboense,Polypodium induens var. subdentatum, Polypodium insidiosum, Polypodiuminsigne, Polypodium intermedium subsp. masafueranum var. obtuseserratum,Polypodium intramarginale, Polypodium involutum, Polypodium itatiayense,Polypodium javanicum, Polypodium juglandifolium, Polypodium kaniense,Polypodium knowltoniorum, Polypodium kyimbilense, Polypodiuml'herminieri var. costaricense, Polypodium lachniferum f. incurvata,Polypodium lachniferum var. glabrescens, Polypodium lachnopus,Polypodium lanceolatum var. cornplanatum, Polypodium lanceolatum var.trichophorum, Polypodium latevagans, Polypodium laxifrons, Polypodiumlaxifrons var. lividum, Polypodium lehmannianum, Polypodium leiorhizum,Polypodium leptopodon, Polypodium leuconeuron var. angustifolia,Polypodium leuconeuron var. latifolium, Polypodium leucosticta,Polypodium limulum, Polypodium lindigii, Polypodium lineatum, Polypodiumlomarioides, Polypodium longifrons, Polypodium loretense, Polypodiumloriceum var. umbraticum, Polypodium loriforme, Polypodium loxogramme f.gigas, Polypodium ludens, Polypodium luzonicum, Polypodium lycopodioidesf. obtusum, Polypodium lycopodioides L., Polypodium macrolepis,Polypodium macrophyllum, Polypodium macrosorum, Polypodiummacrosphaerum, Polypodium maculosum, Polypodium madrense, Polypodiummanmeiense, Polypodium margaritiferum, Polypodium maritimum, Polypodiummartensii, Polypodium mayoris, Polypodium megalolepis, Polypodiummelanotrichum, Polypodium menisciifolium var. pubescens, Polypodiummeniscioides, Polypodium merrillii, Polypodium mettenii, Polypodiummexiae, Polypodium microsorum, Polypodium militare, Polypodium minimum,Polypodium minusculum, Polypodium mixtum, Polypodium mollendense,Polypodium mollissimum, Polypodium moniliforme var. minus, Polypodiummonoides, Polypodium monticola, Polypodium montigenum, Polypodiummoritzianum, Polypodium moultonii, Polypodium multicaudatum, Polypodiummultilineatum, Polypodium multisorum, Polypodium munchii, Polypodiummuscoides, Polypodium myriolepis, Polypodium myriophyllum, Polypodiummyriotrichum, Polypodium nematorhizon, Polypodium nemorale, Polypodiumnesioticum, Polypodium nigrescentium, Polypodium nigripes, Polypodiumnigrocinctum, Polypodium nimbatum, Polypodium nitidissimum, Polypodiumnitidissimum var. latior, Polypodium nubrigenum, Polypodium oligolepis,Polypodium oligosorum, Polypodium oligosorum, Polypodium olivaceum,Polypodium olivaceum var. elatum, Polypodium oodes, Polypodiumoosphaerum, Polypodium oreophilum, Polypodium ornatissimum, Polypodiumornatum, Polypodium ovatum, Polypodium oxylobum, Polypodium oxypholis,Polypodium pakkaense, Polypodium pallidum, Polypodium palmatopedatum,Polypodium palmeri, Polypodium panamense, Polypodium parvum, Polypodiumpatagonicum, Polypodium paucisorum, Polypodium pavonianum, Polypodiumpectinatum var. caliense, Polypodium pectinatum var. hispidum,Polypodium pellucidum, Polypodium pendulum var. boliviense, Polypodiumpercrassum, Polypodium perpusillum, Polypodium peruvianum var.subgibbosum, Polypodium phyllitidis var. elongatum, Polypodiumpichinchense, Polypodium pilosissimum, Polypodium pilosissimum var.glabriusculum, Polypodium pilossimum var. tunguraquensis, Polypodiumpityrolepis, Polypodium platyphyllum, Polypodium playfairii, Polypodiumplebeium var. cooperi, Polypodium plectolepidioides, Polypodiumpleolepis, Polypodium plesiosorum var.i, Polypodium podobasis,Polypodium podocarpum, Polypodium poloense, Polypodium polydatylon,Polypodium polypodioides var. aciculare, Polypodium polypodioides var.michauxianum, Polypodium praetermissum, Polypodium preslianum var.immersum, Polypodium procerum, Polypodium procerum, Polypodiumproductum, Polypodium productum, Polypodium prolongilobum, Polypodiumpropinguum, Polypodium proteus, Polypodium pruinatum, Polypodiumpseudocapillare, Polypodium pseudofratemum, Polypodium pseudonutans,Polypodium pseudoserratum, Polypodium pulcherrimum, Polypodiumpulogense, Polypodium pungens, Polypodium purpusii, Polypodium radicale,Polypodium randallii, Polypodium ratiborii, Polypodium reclinaturn,Polypodium recreense, Polypodium repens var. abruptum, Polypodiumrevolvens, Polypodium rhachipterygium, Polypodium rhomboideum,Polypodium rigens, Polypodium robustum, Polypodium roraimense,Polypodium roraimense, Polypodium rosei, Polypodium rosenstockii,Polypodium rubidum, Polypodium rudimentum, Polypodium rusbyi, Polypodiumsablanianum, Polypodium sarmentosum, Polypodium saxicola, Polypodiumschenckii, Polypodium schlechteri, Polypodium scolopendria, Polypodiumscolopendria, Polypodium scolopendrium, Polypodium scouleri, Polypodiumscutulatum, Polypodium segregatum, Polypodium semihirsutum, Polypodiumsemihirsutum var. fuscosetosum, Polypodium senile var. minor, Polypodiumsericeolanatum, Polypodium serraeforme, Polypodium serricula, Polypodiumsesquipedala, Polypodium sessilifolium, Polypodium setosum var. calvum,Polypodium setulosum, Polypodium shaferi, Polypodium sibomense,Polypodium siccum, Polypodium simacense, Polypodium simulans, Polypodiumsingeri, Polypodium sinicum, Polypodium sintenisii, Polypodium skutchii,Polypodium sloanei, Polypodium sodiroi, Polypodium sordidulum,Polypodium sordidum, Polypodium sphaeropteroides, Polypodium sphenodes,Polypodium sprucei, Polypodium sprucei var. furcativenosa, Polypodiumsteirolepis, Polypodium stenobasis, Polypodium stenolepis, Polypodiumstenopterum, Polypodium subcapillare, Polypodium subflabelliforme,Polypodium subhemionitidium, Polypodium subinaequale, Polypodiumsubintegrum, Polypodium subspathulatum, Polypodium subtile, Polypodiumsubvestitum, Polypodium subviride, Polypodium superficiale var.attenuatum, Polypodium superficiale var. chinensis, Polypodiumsursumcurrens, Polypodium tablazianurn, Polypodium taenifolium,Polypodium tamandarei, Polypodium tatei, Polypodium tenuiculum var.acrosora, Polypodium tenuiculum var. brasiliense, Polypodium tenuilore,Polypodium tenuinerve, Polypodium tepuiense, Polypodium teresae,Polypodium tetragonum var. incompletum, Polypodium thysanolepis var.bipinnatifidum, Polypodium thyssanolepis, var. thyssanolepis, Polypodiumthyssanolepsi, Polypodium tobagense, Polypodium trichophyllum,Polypodium tridactylum, Polypodium tridentatum, Polypodium trifurcaturnvar. brevipes, Polypodium triglossum, Polypodium truncatulum, Polypodiumtruncicola var. major, Polypodium truncicola var. minor, Polypodiumtuberosum, Polypodium tunguraguae, Polypodium turquinurn, Polypodiumturrialbae, Polypodium ursipes, Polypodium vagans, Polypodiumvaldealaturn, Polypodium versteegii, Polypodium villagranii, Polypodiumvirginianum f. cambroideum, Polypodium virginianurn f. peraferens,Polypodium vittarioides, Polypodium vulgare, Polypodium vulgare L.,Polypodium vulgare subsp. oreophilum, Polypodium vulgare var.acuminatum, Polypodium vulpinum, Polypodium williamsii, Polypodiumwobbense, Polypodium×fallacissimum-guttatum, Polypodium xantholepis,Polypodium xiphopteris, Polypodium yarumalense, Polypodium yungense, andPolypodium zosteriforme.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium. In some embodiments the PtIP-50 polypeptide is derived froma fern species in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium selected from but not limited to Platycerium alcicorne,Platycerium andinum, Platycerium angolense, Platycerium bifurcatum,Platycerium coronarium, Platycerium elephantotis, Platycerium ellisfi,Platycerium grande, Platycerium hillii, Platycerium holttumfi,Platycerium madagascariense, Platycerium quadridichotomum, Platyceriumridleyi, Platycerium sp. ES-2011, Platycerium stemaria, Platyceriumsuperbum, Platycerium veitchii, Platycerium wallichii, Platyceriumwandae, Platycerium wilhelminae-reginae, and Platycerium willinckii.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Division Lycophyta.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Class Isoetopsida or Class Lycopodiopsida.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Class Isoetopsida Order Selaginales. In some embodiments the PtIP-50polypeptide is derived from a fern species in the Class Isoetopsida,Order Selaginales, Family Selaginellaceae. In some embodiments thePtIP-50 polypeptide is derived from a species in the Genus Selaginella.In some embodiments the PtIP-50 polypeptide is derived from aSelaginella species selected from but not limited to Selaginellaacanthonota, Selaginella apoda, Selaginella arbuscula, Selaginellaarenicola, Selaginella arizonica, Selaginella armata, Selaginellaasprella, Selaginella biformis, Selaginella bigelovii, Selaginellabraunii, Selaginella cinerascens, Selaginella cordifolia, Selaginelladeflexa, Selaginella delicatula, Selaginella densa, Selaginelladouglasii, Selaginella eatonii, Selaginella eclipes, Selaginellaeremophila, Selaginella erythropus, Selaginella flabellata, Selaginellahansenii, Selaginella heterodonta, Selaginella kraussiana, Selaginellakrugii, Selaginella laxifolia, Selaginella lepidophylla, Selaginellaleucobryoides, Selaginella ludoviciana, Selaginella mutica, Selaginellaoregana, Selaginella ovifolia, Selaginella pallescens, Selaginellaperuviana, Selaginella pilifera, Selaginella plana, Selaginella plumosa,Selaginella pulcherrima, Selaginella rupestris, Selaginella rupincola,Selaginella scopulorum, Selaginella selaginoides, Selaginella sibirica,Selaginella standleyi, Selaginella stellata, Selaginella subcaulescens,Selaginella substipitata, Selaginella tenella, Selaginella tortipila,Selaginella uliginosa, Selaginella umbrosa, Selaginella uncinata,Selaginella underwoodii, Selaginella utahensis, Selaginella victoriae,Selaginella viridissima, Selaginella wallacei, Selaginella watsonii,Selaginella weatherbiana, Selaginella willdenowii, Selaginella wrightiiand Selaginella×neomexicana.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Class Lycopodiopsida, Order Lycopodiales.

In some embodiments the PtIP-50 polypeptide is derived from a fernspecies in the Class Lycopodiopsida, Order Lycopodiales FamilyLycopodiaceae or Family Huperziaceae.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Genus Austrolycopodium, Dendrolycopodium, Diphasiastrum, Diphasium,Huperzia, Lateristachys, Lycopodiastrum, Lycopodiella, Lycopodium,Palhinhaea, Pseudodiphasium, Pseudolycopodiella, Pseudolycopodium orSpinulum.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Genus Lycopodium.

In some embodiments the PtIP-50 polypeptide is derived from a Lycopodiumspecies selected from but not limited to Lycopodium alpinum L.,Lycopodium annotinum L., Lycopodium clavatum L., Lycopodium cornplanatumL., Lycopodium dendroideum Michx., Lycopodium digitatum, Lycopodiumxhabereri, Lycopodium hickeyi, Lycopodium×issleri, Lycopodium lagopus,Lycopodium obscurum L., Lycopodium phlegmaria L., Lycopodiumsabinifolium, Lycopodium sitchense, Lycopodium tristachyum, Lycopodiumvenustulum, Lycopodium venustulum var. venustulum, Lycopodium venustulumvar. verticale, Lycopodium volubile and Lycopodium×zeilleri.

In some embodiments the PtIP-50 polypeptide is derived from a species inthe Genus Huperzia. In some embodiments the PtIP-50 polypeptide isderived from a species selected from but not limited to Huperziaappressa, Huperzia arctica, Huperzia attenuata, Huperzia australiana,Huperzia balansae, Huperzia billardierei, Huperzia brassii, Huperziacampiana, Huperzia capellae, Huperzia carinata, Huperzia cf. carinataARF000603, Huperzia cf. nummulariifolia ARF001140, Huperzia cf.phlegmaria ARF000717, Huperzia cf. phlegmaria ARF000771, Huperzia cf.phlegmaria ARF000785, Huperzia cf. phlegmaria ARF001007, Huperzia cf.phlegmaria ARF002568, Huperzia cf. phlegmaria ARF002703, Huperzia cf.phlegmaria Wikstrom 1998, Huperzia chinensis, Huperzia compacta,Huperzia crassa, Huperzia crispata, Huperzia cryptomeriana, Huperziacumingii, Huperzia dacrydioides, Huperzia dalhousieana, Huperziadichotoma, Huperzia emeiensis, Huperzia ericifolia, Huperzia eversa,Huperzia fargesii, Huperzia fordii, Huperzia funiformis, Huperziagoebellii, Huperzia haleakalae, Huperzia hamiltonii, Huperziaheteroclita, Huperzia hippuridea, Huperzia hippuris, Huperzia holstii,Huperzia horizontalis, Huperzia hunanensis, Huperzia hystrix, Huperzialindenii, Huperzia linifolia, Huperzia lockyeri, Huperzia lucidula,Huperzia mingcheensis, Huperzia miyoshiana, Huperzia nanchuanensis,Huperzia nummulariifolia, Huperzia obtusifolia, Huperziaophioglossoides, Huperzia petiolata, Huperzia phlegmaria, Huperziaphlegmarioides, Huperzia phyllantha, Huperzia pinifolia, Huperziapolydactyla, Huperzia prolifera, Huperzia reflexa, Huperziarosenstockiana, Huperzia rufescens, Huperzia salvinoides, Huperziasarmentosa, Huperzia selago, Huperzia serrata, Huperzia sieboldii,Huperzia somae, Huperzia squarrosa, Huperzia subulata, Huperziasutchueniana, Huperzia tauri, Huperzia taxifolia, Huperzia tenuis,Huperzia tetragona, Huperzia tetrasticha, Huperzia unguiculata, Huperziavaria, Huperzia verticillata and Huperzia wilsonii.

Phylogenetic, Sequence Motif, and Structural Analyses for InsecticidalProtein Families

The sequence and structure analysis method employed is composed of fourcomponents: phylogenetic tree construction, protein sequence motifsfinding, secondary structure prediction, and alignment of proteinsequences and secondary structures. Details about each component areillustrated below.

1) Phylogenetic Tree Construction

The phylogenetic analysis was performed using the software MEGA5.Protein sequences were subjected to ClustalW version 2 analysis (LarkinM. A et al (2007) Bioinformatics 23(21): 2947-2948) for multiplesequence alignment. The evolutionary history was then inferred by theMaximum Likelihood method based on the JTT matrix-based model. The treewith the highest log likelihood was obtained, exported in Newick format,and further processed to extract the sequence IDs in the same order asthey appeared in the tree. A few clades representing sub-families weremanually identified for each insecticidal protein family.

2) Protein Sequence Motifs Finding

Protein sequences were re-ordered according to the phylogenetic treebuilt previously, and fed to the motif analysis tool MEME (Multiple EMfor motif Elicitation) (Bailey T. L., and Elkan C., Proceedings of theSecond International Conference on Intelligent Systems for MolecularBiology, pp. 28-36, AAAI Press, Menlo Park, Calif., 1994) foridentification of key sequence motifs. MEME was setup as follows:Minimum number of sites 2, Minimum motif width 5, and Maximum number ofmotifs 50. Sequence motifs unique to each sub-family were identified byvisual observation. The distribution of motifs across the entire genefamily could be visualized in HTML webpage. The motifs are numberedrelative to the ranking of the E-value for each motif. The amino acidsequence motifs identified for each of the PtIP-50 polypeptides and theresidue ranges defining the motifs relative to each of the correspondingsequence identifier (SEQ ID NO:) are shown in Table 2. An amino acidsequence motif not identified in a particular PtIP-50 polypeptide isindicated in Table 2 as “n. i.”. The amino acid sequence motifsidentified for each of the PtIP-65 polypeptides and the residue rangesdefining the motifs relative to the corresponding sequence identifier(SEQ ID NO:) are shown in Table 3. An amino acid sequence motif notidentified in a particular PtIP-65 polypeptide is indicated in Table 3as “n. i.”. FIG. 16a -16u shows an alignment of the PtIP-50 polypeptidesPtIP-50Aa (SEQ ID NO: 71), PtIP-50Ba (SEQ ID NO: 77), PtIP-50Bb (SEQ IDNO: 84), PtIP-50Bc (SEQ ID NO: 78), PtIP-50Bd (SEQ ID NO: 96), PtIP-50Fa(SEQ ID NO: 86), PtIP-50Fb (SEQ ID NO: 72), PtIP-50Fd (SEQ ID NO: 87),PtIP-50Fe (SEQ ID NO: 79), PtIP-50Ff (SEQ ID NO: 83), PtIP-50Fg (SEQ IDNO: 82), PtIP-50Fh (SEQ ID NO: 81), PtIP-50Fi (SEQ ID NO: 90), PtIP-50Fj(SEQ ID NO: 91), PtIP-50Fk (SEQ ID NO: 89), PtIP-50Fl (SEQ ID NO: 94),PtIP-50Fm (SEQ ID NO: 98), PtIP-50Fn (SEQ ID NO: 93), PtIP-50Fo (SEQ IDNO: 73), PtIP-50Fp (SEQ ID NO: 97), PtIP-50Fq (SEQ ID NO: 95), PtIP-50Fr(SEQ ID NO: 75), PtIP-50Fs (SEQ ID NO: 74), PtIP-50Ft (SEQ ID NO: 85),PtIP-50Ga (SEQ ID NO: 88), PtIP-50Gb (SEQ ID NO: 80, PtIP-50Gc (SEQ IDNO: 76), and PtIP-50Gd (SEQ ID NO: 92)], and the location, relative toPtIP-50Fb (SEQ ID NO: 72), of the amino acid sequence motifs present inPtIP-50Fb (SEQ ID NO: 72). FIGS. 17a-17k shows an alignment of the aminoacid sequences of PtIP-65Aa (SEQ ID NO: 22), PtIP-65Ba (SEQ ID NO: 24),PtIP-65Bb (SEQ ID NO: 26), PtIP-65Ca (SEQ ID NO: 29), PtIP-65Fa (SEQ IDNO: 41), PtIP-65Fb (SEQ ID NO: 42), PtIP-65Ga (SEQ ID NO: 30), PtIP-65Gb(SEQ ID NO: 34), PtIP-65Gc (SEQ ID NO: 27), PtIP-65Gd (SEQ ID NO: 35),PtIP-65Ge (SEQ ID NO: 36), PtIP-65Ha (SEQ ID NO: 25), PtIP-65Hb (SEQ IDNO: 31), PtIP-65Hc (SEQ ID NO: 23), PtIP-65Hd (SEQ ID NO: 28), PtIP-65He(SEQ ID NO: 32), PtIP-65Hf (SEQ ID NO: 33), PtIP-65Hg (SEQ ID NO: 40),PtIP-65Hh (SEQ ID NO: 38), PtIP-65Hj (SEQ ID NO: 39), and PtIP-65Hk (SEQID NO: 37); and the location, relative to PtIP-65Gc (SEQ ID NO: 27), ofthe amino acid sequence motifs present in PtIP-65Gc (SEQ ID NO: 72).

3) Secondary Structure Prediction

PSIPRED, top ranked secondary structure prediction method (Jones D T.(1999) J. Mol. Biol. 292: 195-202), was installed in local Linux server,and used for protein secondary structure prediction. The tool providesaccurate structure prediction using two feed-forward neural networksbased on the PSI-BLAST output. The PSI-BLAST database was created byremoving low-complexity, transmembrane, and coiled-coil regions inUniref100. The PSIPRED results contain the PtIP-secondary structures(Alpha helix: H, Beta strand: E, and Coil: C) and the correspondingconfidence scores for each amino acid in a given protein sequence. FIG.16 shows the PtIP-50 polypeptide amino acid sequence alignments and theconserved secondary structural regions. FIG. 17 shows the PtIP-65polypeptide amino acid sequence alignments and conserved secondarystructural regions.

4) Alignment of Protein Sequences and Secondary Structures

A customized script was developed to generate gapped secondary structurealignment according to the multiple protein sequence alignment from step1 for all proteins. All aligned protein sequences and structures wereconcatenated into a single FASTA file, and then imported into MEGA forvisualization and identification of conserved structures. The file wasalso edited in GeneDoc to produce acceptable sequence format for patentoffices.

In some embodiments the PtIP-50 polypeptide comprises at least one aminoacid sequence motif as shown in Table 2.

TABLE 2 Motif 23 Motif 8 Motif 16 Motif 18 Motif 14 Motif 19 PtIP-50AaSEQ ID NO: 71 5-16 17-44 49-70 71-91 196-226 231-252 PtIP-50Ba SEQ IDNO: 77 5-16 17-44 49-70 71-91 196-226 231-252 PtIP-50Bb SEQ ID NO: 845-16 17-44 49-70 71-91 195-225 230-251 PtIP-50Bc SEQ ID NO: 78 5-1617-44 49-70 71-91 195-225 230-251 PtIP-50Bd SEQ ID NO: 96 5-16 17-4449-70 71-91 195-225 230-251 PtIP-50Fa SEQ ID NO: 86 2-13 14-41 48-6973-93 253-283 289-310 PtIP-50Fb SEQ ID NO: 72 7-18 21-48 49-70 73-93235-265 270-291 PtIP-50Fd SEQ ID NO: 87 12-23  26-53 54-75  86-106277-307 312-333 PtIP-50Fe SEQ ID NO: 79 7-18 21-48 49-70 73-93 233-263268-289 PtIP-50Ff SEQ ID NO: 83 2-13 14-41 42-63 70-90 264-294 299-320PtIP-50Fg SEQ ID NO: 82 11-22  25-52 53-74 76-96 288-318 323-344PtIP-50Fh SEQ ID NO: 81 11-22  25-52 53-74  82-102 285-315 320-341PtIP-50Fi SEQ ID NO: 90 2-13 14-41 n.i. 63-83 237-267 272-293 PtIP-50FjSEQ ID NO: 91 2-13 14-41 48-69 72-92 262-292 297-318 PtIP-50Fk SEQ IDNO: 89 54-65  68-95  96-117 120-140 281-311 316-337 PtIP-50Fl SEQ ID NO:94 7-18 21-48 49-70 73-93 234-264 269-290 PtIP-50Fm SEQ ID NO: 98 17-28 31-58 59-80  91-111 283-313 318-339 PtIP-50Fn SEQ ID NO: 93 7-18 21-4849-70 73-93 236-266 271-292 PtIP-50Fo SEQ ID NO: 73 50-61  64-91  92-113124-144 285-315 320-341 PtIP-50Fp SEQ ID NO: 97 6-17 18-45 46-67 68-88223-253 259-280 PtIP-50Fq SEQ ID NO: 95 n.i. n.i. 33-54 61-81 257-287292-313 PtIP-50Fr SEQ ID NO: 75 11-22  25-52 53-74  85-105 278-308313-334 PtIP-50Fs SEQ ID NO: 74 7-18 21-48 49-70  80-100 242-272 277-298PtIP-50Ft SEQ ID NO: 85 7-18 21-48 49-70 79-99 265-295 300-321 PtIP-50GaSEQ ID NO: 88 40-51  54-81  82-103 106-126 256-286 292-313 PtIP-50Gb SEQID NO: 80 39-50  53-80  81-102 105-125 256-286 292-313 PtIP-50Gc SEQ IDNO: 76 40-51  54-81  82-103 106-126 256-286 292-313 PtIP-50Gd SEQ ID NO:92 41-52  55-82  83-104 107-127 258-288 294-315 Motif 20 Motif 13 Motif2 motif 12 Motif 11 Motif 21 PtIP-50Aa SEQ ID NO: 71 255-270 273-294295-337 342-377 385-410 412-427 PtIP-50Ba SEQ ID NO: 77 255-270 273-294295-337 342-377 385-410 412-427 PtIP-50Bb SEQ ID NO: 84 254-269 272-293294-336 341-376 384-409 411-426 PtIP-50Bc SEQ ID NO: 78 254-269 272-293294-336 341-376 384-409 411-426 PtIP-50Bd SEQ ID NO: 96 254-269 272-293294-336 341-376 384-409 411-426 PtIP-50Fa SEQ ID NO: 86 311-326 330-351352-394 399-434 442-467 472-487 PtIP-50Fb SEQ ID NO: 72 298-313 318-339340-382 387-422 430-455 457-472 PtIP-50Fd SEQ ID NO: 87 339-354 359-389381-423 428-463 471-496 498-513 PtIP-50Fe SEQ ID NO: 79 296-311 316-337338-380 385-420 428-453 455-470 PtIP-50Ff SEQ ID NO: 83 321-336 340-361362-404 409-444 452-477 479-494 PtIP-50Fg SEQ ID NO: 82 350-365 370-391392-434 439-474 482-507 509-524 PtIP-50Fh SEQ ID NO: 81 348-363 368-389393-435 440-475 483-508 510-525 PtIP-50Fi SEQ ID NO: 90 297-312 316-337338-380 385-420 428-453 455-470 PtIP-50Fj SEQ ID NO: 91 330-345 349-370371-413 418-453 461-486 488-503 PtIP-50Fk SEQ ID NO: 89 344-359 364-385386-428 433-468 476-501 503-518 PtIP-50Fl SEQ ID NO: 94 297-312 317-338339-381 386-421 429-454 456-471 PtIP-50Fm SEQ ID NO: 98 345-360 365-386387-429 434-469 477-502 504-519 PtIP-50Fn SEQ ID NO: 93 299-314 319-340341-383 388-423 431-456 458-473 PtIP-50Fo SEQ ID NO: 73 348-363 368-389390-432 437-472 480-50  507-522 PtIP-50Fp SEQ ID NO: 97 288-303 309-330331-373 378-413 421-446 n.i. PtIP-50Fq SEQ ID NO: 95 314-329 337-358359-401 406-441 449-474 476-491 PtIP-50Fr SEQ ID NO: 75 340-355 360-381382-424 429-464 473-498 500-515 PtIP-50Fs SEQ ID NO: 74 305-320 325-346347-389 394-429 437-462 464-479 PtIP-50Ft SEQ ID NO: 85 329-344 350-371372-414 419-454 462-487 489-504 PtIP-50Ga SEQ ID NO: 88 320-335 341-362363-405 410-445 453-478 n.i. PtIP-50Gb SEQ ID NO: 80 320-335 341-362363-405 410-445 453-478 n.i. PtIP-50Gc SEQ ID NO: 76 320-335 341-362363-405 410-445 453-478 n.i. PtIP-50Gd SEQ ID NO: 92 322-337 343-364365-407 412-447 455-480 n.i. Motif 9 Motif 6 Motif 1 Motif 4 Motif 7Motif 22 PtIP-50Aa SEQ ID NO: 71 429-478 480-501 504-553 556-598 606-652653-664 PtIP-50Ba SEQ ID NO: 77 429-478 480-501 504-553 556-598 606-652653-664 PtIP-50Bb SEQ ID NO: 84 428-477 479-500 503-552 555-597 605-651652-663 PtIP-50Bc SEQ ID NO: 78 428-477 479-500 503-552 555-597 605-651652-663 PtIP-50Bd SEQ ID NO: 96 428-477 479-500 503-552 555-597 605-651652-663 PtIP-50Fa SEQ ID NO: 86 489-538 540-561 564-613 616-658 n.i.718-729 PtIP-50Fb SEQ ID NO: 72 474-523 525-546 549-598 601-643 651-697698-709 PtIP-50Fd SEQ ID NO: 87 515-564 566-587 590-639 642-684 692-738739-750 PtIP-50Fe SEQ ID NO: 79 472-521 523-544 547-596 599-641 649-695696-707 PtIP-50Ff SEQ ID NO: 83 496-545 547-568 571-620 623-665 674-720721-732 PtIP-50Fg SEQ ID NO: 82 526-575 577-598 601-650 653-695 703-749750-761 PtIP-50Fh SEQ ID NO: 81 527-576 578-599 602-651 655-697 705-751752-763 PtIP-50Fi SEQ ID NO: 90 472-521 523-544 548-597 600-642 651-697698-709 PtIP-50Fj SEQ ID NO: 91 505-554 556-577 581-630 633-675 684-730731-742 PtIP-50Fk SEQ ID NO: 89 520-569 571-592 595-644 647-689 697-743744-755 PtIP-50Fl SEQ ID NO: 94 473-522 524-545 548-597 600-642 650-696697-708 PtIP-50Fm SEQ ID NO: 98 521-570 572-593 596-645 648-690 698-744745-756 PtIP-50Fn SEQ ID NO: 93 475-524 526-547 550-599 602-644 652-698699-710 PtIP-50Fo SEQ ID NO: 73 524-573 575-596 599-648 651-693 701-747748-759 PtIP-50Fp SEQ ID NO: 97 465-514 516-537 540-589 592-634 641-687688-699 PtIP-50Fq SEQ ID NO: 95 493-542 544-565 568-617 620-662 671-717718-729 PtIP-50Fr SEQ ID NO: 75 517-566 568-589 592-641 644-686 694-740741-752 PtIP-50Fs SEQ ID NO: 74 481-530 532-553 556-605 608-650 658-704705-716 PtIP-50Ft SEQ ID NO: 85 506-555 557-578 581-630 633-675 683-729730-741 PtIP-50Ga SEQ ID NO: 88 504-553 554-575 578-627 631-673 n.i.732-743 PtIP-50Gb SEQ ID NO: 80 504-553 554-575 578-627 631-673 n.i.732-743 PtIP-50Gc SEQ ID NO: 76 n.i. 554-575 578-627 631-673 n.i.732-743 PtIP-50Gd SEQ ID NO: 92 n.i. 556-577 580-629 633-675 n.i.734-745 Motif 10 Motif 17 Motif 3 Motif 24 Motif 5 Motif 15 PtIP-50AaSEQ ID NO: 71 665-686 690-705 709-746 749-757 758-779 781-802 PtIP-50BaSEQ ID NO: 77 665-686 690-705 709-746 749-757 758-779 781-802 PtIP-50BbSEQ ID NO: 84 664-685 691-706 710-747 750-758 759-780 782-803 PtIP-50BcSEQ ID NO: 78 664-685 689-704 708-745 748-756 757-778 780-801 PtIP-50BdSEQ ID NO: 96 664-685 689-704 708-745 748-756 757-778 780-801 PtIP-50FaSEQ ID NO: 86 731-752 763-778 780-817 820-828 829-850 853-874 PtIP-50FbSEQ ID NO: 72 711-732 747-762 764-801 804-812 813-834 837-858 PtIP-50FdSEQ ID NO: 87 752-773 784-799 801-838 841-849 850-871 873-894 PtIP-50FeSEQ ID NO: 79 709-730 740-755 757-794 797-805 806-828 830-851 PtIP-50FfSEQ ID NO: 83 734-755 766-781 783-820 823-831 832-853 855-876 PtIP-50FgSEQ ID NO: 82 763-784 789-804 806-843 846-854 855-876 878-899 PtIP-50FhSEQ ID NO: 81 765-786 794-809 811-848 851-859 860-881 883-904 PtIP-50FiSEQ ID NO: 90 712-733 743-758 760-797 800-808 809-830 832-853 PtIP-50FjSEQ ID NO: 91 744-765 775-790 792-829 832-840 842-863 865-886 PtIP-50FkSEQ ID NO: 89 757-778 792-807 809-846 849-857 858-879 882-903 PtIP-50FlSEQ ID NO: 94 710-731 742-757 759-796 799-807 808-829 832-853 PtIP-50FmSEQ ID NO: 98 758-779 786-801 803-840 843-851 852-873 875-896 PtIP-50FnSEQ ID NO: 93 712-733 744-759 761-798 801-809 810-831 834-855 PtIP-50FoSEQ ID NO: 73 761-782 791-806 808-845 848-856 857-878 881-902 PtIP-50FpSEQ ID NO: 97 701-722 727-742 744-781 784-792 793-814 819-839 PtIP-50FqSEQ ID NO: 95 731-752 763-778 780-817 820-828 830-851 853-874 PtIP-50FrSEQ ID NO: 75 754-775 789-804 806-843 846-854 855-876 879-900 PtIP-50FsSEQ ID NO: 74 718-739 752-767 769-806 809-817 818-839 842-863 PtIP-50FtSEQ ID NO: 85 743-764 769-784 786-823 826-834 835-856 858-879 PtIP-50GaSEQ ID NO: 88 746-767 769-784 786-823 824-832 833-854 860-881 PtIP-50GbSEQ ID NO: 80 746-767 769-784 786-823 824-832 833-854 859-880 PtIP-50GcSEQ ID NO: 76 746-767 769-784 786-823 824-832 833-854 860-881 PtIP-50GdSEQ ID NO: 92 748-769 771-786 788-825 826-834 835-856 858-879

In some embodiments the PtIP-50 polypeptide comprises at least one aminoacid sequence motif selected from: an amino acid sequence motif 1 asrepresented by an amino acid sequence of the formulaAWK[AT][KS]C[KR]NVA[AV][LV]G[RQ]E[ML][CTS]TTAA[YH]I[SA][EQ]LQY[DEQ]I[QK]VQ[AE]MLQ[EDQ]IA[RQ][KR]QA[DE]RL[SE][SGA]I(SEQ ID NO: 123); an amino acid sequence motif 2 as represented by anamino acid sequence of the formula[EQ][SA]YDQ[ESD][LF][KR]Q[LF][KQ]LF[IV][AQ]QN[KE]ILG[SG]YLL[EQ]QN[RK]AFA[EA][KR]E[RK]DM[ED][VA]FHS(SEQ ID NO: 124); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaF[HR]DRN[EK][TASGERDN]E[VP]L[HE]YEA[AG][TV][PG]L[DV]Y[HQ]YAY[NR]LDTG[EA]TT[LV][TS]NLPS(SEQ ID NO: 125); an amino acid sequence motif 4 as represented by anamino acid sequence of the formulaADL[ST][SN][YF]TE[ML][VA][TS][QE]MDMRTTR[LM]L[LV][EA]LIKVL[HNY][IM]QN[AGV]ALMY[QE]YLS[EP][PA] SEQ ID NO: 126); an amino acid sequence motif 5 asrepresented by an amino acid sequence of the formulaPFT[RNT]WRLR[LV]SASA[EQ]EN[EKQ]GLAFP (SEQ ID NO: 127); an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaDW[LD]IF[VE]NE[VI]E[AG]VA[EA]QMP[TE]EVSE (SEQ ID NO: 128); an amino acidsequence motif 7 as represented by an amino acid sequence of the formulaV[TN]M[EDQ]TVW[GR][ML]L[VI]Q[HQ]E[HQ]AA[IV][LQ]GL[LM][RQ]LG[PA][SAP][FS]D[FI][TRV]RT[YF]VVKDIPV[DS]LLL[DH]G (SEQ ID NO: 129); an amino acid sequencemotif 8 as represented by an amino acid sequence of the formulaIRLD[QK]MEFSEVM[VA][IV]HRM[FH][IVF][RK][LM][DP][DE]LD[IVL][AG][HL] (SEQID NO: 130); an amino acid sequence motif 9 as represented by an aminoacid sequence of the formula[AL]A[RE][GK]L[QKE][RK]V[LV][EQ]IL[ED][GD]LQA[VA]MEVV[AK][AI]I[KN][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][ML]P[TS]E (SEQ ID NO: 131); an amino acidsequence motif 10 as represented by an amino acid sequence of theformula [TV]FP[LS][TS][WL]SRVRI[HR][HY][LV]EMKF[VD][GQA][AG]A (SEQ IDNO: 132); an amino acid sequence motif 11 as represented by an aminoacid sequence of the formula[RK]Q[VL]A[RN]A[LV]FAV[LF][GR]AI[AG][AS][VI][AG]L[AT][FVL][ALV]TGGA (SEQID NO: 133); an amino acid sequence motif 12 as represented by an aminoacid sequence of the formula[QL][RK][QRT][STI]EL[DQ]N[TA][IML][QE][KR][MI]DQLSLQMET[QE]S[AE][DA]M[ED]QA[KQR][AE]DM[ED][AE] (SEQ ID NO: 134); an amino acid sequence motif 13 asrepresented by an amino acid sequence of the formulaLVVP[QR]LQY[DH]MYSNLI[ND][RQ][ML][AV][QR]VA (SEQ ID NO: 135); an aminoacid sequence motif 14 as represented by an amino acid sequence of theformula[PA][DV]A[LV]LTDP[NS][IV][LI][LS][GC]MQT[ST][ML]LIAELV[EL][FV][AS][HQ][PN]SS(SEQ ID NO: 136); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[AT][TV][SG]A[DG][AD]TTQIAITF[HYF][VI][ST]AIR[RQE]I (SEQ ID NO: 137); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula LEGA[ED]KVKRLYVFADVVEL[PE][SG]K (SEQ ID NO: 138); an aminoacid sequence motif 17 as represented by an amino acid sequence of theformula P[IV]TD[TS]GE[VI]Y[IM]LLQ[SG]SR (SEQ ID NO: 139); an amino acidsequence motif 18 as represented by an amino acid sequence of theformula T[VL][RS]LPG[ST][IV][MS]VVILCR[VI]L[HV][LV]N[GS] (SEQ ID NO:140); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula [AV]V[TR][KR]H[VA]EWLN[KT]LL[LV]Q[AV]SAAAQ[GS](SEQ ID NO: 141); an amino acid sequence motif 20 as represented by anamino acid sequence of the formula[YV][LV]ALL[FY]R[AT]Q[YA][LVF][IL]K[LGM]V[GK] (SEQ ID NO: 142); an aminoacid sequence motif 21 as represented by an amino acid sequence of theformula AP[GAL]A[VIM][AS]AA[KGR][GA]AV[ST][AI]AG (SEQ ID NO: 143); anamino acid sequence motif 22 as represented by an amino acid sequence ofthe formula [ED]D[WY][EQ]FE[IL]PVEDF (SEQ ID NO: 144); and an amino acidsequence motif 23 as represented by an amino acid sequence of theformula YS[EH]LYR[DE][LV]NQ[VI]S (SEQ ID NO: 145); and an amino acidsequence motif 24 as represented by an amino acid sequence of theformula F[AI][NR]TFMRMT (SEQ ID NO: 146).

In some embodiments the PtIP-50 polypeptide comprises at least one aminoacid sequence motif selected from: an amino acid sequence motif 1 havingat least 90% identity to an amino acid sequence ofAWK[AT][KS]C[KR]NVA[AV][LV]G[RQ]E[ML][CTS]TTAA[YH]I[SA][EQ]LQY[DEQ]I[QK]VQ[AE]MLQ[EDQ]IA[RQ][KR]QA[DE]RL[SE][SGA]I(SEQ ID NO: 123); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of[EQ][SA]YDQ[ESD][LF][KR]Q[LF][KQ]LF[IV][AQ]QN[KE]ILG[SG]YLL[EQ]QN[RK]AFA[EA][KR]E[RK]DM[ED][VA]FHS(SEQ ID NO: 124); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence ofF[HR]DRN[EK][TASGERDN]E[VP]L[HE]YEA[AG][TV][PG]L[DV]Y[HQ]YAY[NR]LDTG[EA]TT[LV][TS]NLPS(SEQ ID NO: 125); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence ofADL[ST][SN][YF]TE[ML][VA][TS][QE]MDMRTTR[LM]L[LV][EA]LIKVL[HNY][IM]QN[AGV]ALMY[QE]YLS[EP][PA] (SEQ ID NO: 126); an amino acid sequence motif 5 havingat least 90% identity to an amino acid sequence ofPFT[RNT]WRLR[LV]SASA[EQ]EN[EKQ]GLAFP (SEQ ID NO: 127); an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof DW[LD]IF[VE]NE[VI]E[AG]VA[EA]QMP[TE]EVSE (SEQ ID NO: 128); an aminoacid sequence motif 7 having at least 90% identity to an amino acidsequence ofV[TN]M[EDQ]TVW[GR][ML]L[VI]Q[HQ]E[HQ]AA[IV][LQ]GL[LM][RQ]LG[PA][SAP][FS]D[FI][TRV]RT[YF]VVKDIPV[DS]LLL[DH]G (SEQ ID NO: 129); an amino acid sequencemotif 8 having at least 90% identity to an amino acid sequence ofIRLD[QK]MEFSEVM[VA][IV]HRM[FH][IVF][RK][LM][DP][DE]LD[IVL][AG][HL] (SEQID NO: 130); an amino acid sequence motif 9 having at least 90% identityto an amino acid sequence of[AL]A[RE][GK]L[QKE][RK]V[LV][EQ]IL[ED][GD]LQA[VA]MEVV[AK][AI]I[KN][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][ML]P[TS]E (SEQ ID NO: 131); an amino acidsequence motif 10 having at least 90% identity to an amino acid sequenceof [TV]FP[LS][TS][WL]SRVRI[HR][HY][LV]EMKF[VD][GQA][AG]A (SEQ ID NO:132); an amino acid sequence motif 11 having at least 90% identity to anamino acid sequence of[RK]Q[VL]A[RN]A[LV]FAV[LF][GR]AI[AG][AS][VI][AG]L[AT][FVL][ALV]TGGA (SEQID NO: 133); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of[QL][RK][QRT][STI]EL[DQ]N[TA][IML][QE][KR][MI]DQLSLQMET[QE]S[AE][DA]M[ED]QA[KQR][AE]DM[ED][AE] (SEQ ID NO: 134); an amino acid sequence motif 13having at least 90% identity to an amino acid sequence ofLVVP[QR]LQY[DH]MYSNLI[ND][RQ][ML][AV][QR]VA (SEQ ID NO: 135); an aminoacid sequence motif 14 having at least 90% identity to an amino acidsequence of[PA][DV]A[LV]LTDP[NS][IV][LI][LS][GC]MQT[ST][ML]LIAELV[EL][FV][AS][HQ][PN]SS(SEQ ID NO: 136); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of[AT][TV][SG]A[DG][AD]TTQIAITF[HYF][VI][ST]AIR[RQE]I (SEQ ID NO: 137); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of LEGA[ED]KVKRLYVFADVVEL[PE][SG]K (SEQ ID NO: 138); anamino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of P[IV]TD[TS]GE[VI]Y[IM]LLQ[SG]SR (SEQ ID NO: 139); anamino acid sequence motif 18 having at least 90% identity to an aminoacid sequence of T[VL][RS]LPG[ST][IV][MS]VVILCR[VI]L[HV][LV]N[GS] (SEQID NO: 140); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of[AV]V[TR][KR]H[VA]EWLN[KT]LL[LV]Q[AV]SAAAQ[GS] (SEQ ID NO: 141); anamino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of [YV][LV]ALL[FY]R[AT]Q[YA][LVF][IL]K[LGM]V[GK] (SEQ IDNO: 142); an amino acid sequence motif 21 having at least 90% identityto an amino acid sequence of AP[GAL]A[VIM][AS]AA[KGR][GA]AV[ST][AI]AG(SEQ ID NO: 143) an amino acid sequence motif 22 having at least 90%identity to an amino acid sequence of [ED]D[WY][EQ]FE[IL]PVEDF (SEQ IDNO: 144); an amino acid sequence motif 23 having at least 90% identityto an amino acid sequence of YS[EH]LYR[DE][LV]NQ[VI]S (SEQ ID NO: 145);an amino acid sequence motif 24 having at least 90% identity to an aminoacid sequence of F[AI][NR]TFMRMT (SEQ ID NO: 146).

In some embodiments the PtIP-50 polypeptide comprises at least one aminoacid sequence motif selected from: an amino acid sequence motif 1 asrepresented by an amino acid sequence of the formulaAWK[ATS][KSRT]C[KR]NVA[AVLI][LVI]G[RQKN]E[MLIV][CTS]TTAA[YHWF]I[SAT][EQDN]LQY[DEQN]I[QKNR]VQ[AED]MLQ[EDQN]IA[RQKN][KR]QA[DE]RL[SETD][SGAT]I (SEQ IDNO: 99); an amino acid sequence motif 2 as represented by an amino acidsequence of the formula[EQND][SAT]YDQ[ESDT][LFIV][KR]Q[LFIV][KQRN]LF[IVL][AQN]QN[KERD]ILG[SGT]YLL[EQDN]QN[RK]AFA[EAD][KR]E[RK]DM[ED][VAIL]FHS (SEQ ID NO: 100); an aminoacid sequence motif 3 as represented by an amino acid sequence of theformulaF[HRK]DRN[EKDR][TASGERDNKQ]E[VPIL]L[HERKD]YEA[AG][TVSIL][PG]L[DVELI]Y[HQN]YAY[NRQK]LDTG[EAD]TT[LVI][TS]NLPS(SEQ ID NO: 101); an amino acid sequence motif 4 as represented by anamino acid sequence of the formulaADL[ST][SNTQ][YFW]TE[MLIV][VAIL][TS][QEDN]MDMRTTR[LMIV]L[LVI][EAD]LIKVL[HNYWF][IMVL]QN[AGVIL]ALMY[QEDN]YLS[EPD][PA] SEQ ID NO: 102); an amino acidsequence motif 5 as represented by an amino acid sequence of the formulaPFT[RNTKQS]WRLR[LVI]SASA[EQDN]EN[EKQDRN]GLAFP (SEQ ID NO: 103); an aminoacid sequence motif 6 as represented by an amino acid sequence of theformula DW[LDIVE]IF[VEILD]NE[VIL]E[AG]VA[EAD]QMP[TESD]EVSE (SEQ ID NO:104); an amino acid sequence motif 7 as represented by an amino acidsequence of the formulaV[TNSQ]M[EDQN]TVW[GRK][MLIV]L[VIL]Q[HQN]E[HQN]AA[IVL][LQIVN]GL[LMIV][RQKN]LG[PA][SAPT][FSWYT]D[FILV][TRVSKIL]RT[YFW]VVKDIPV[DSET]LLL[DHE]G (SEQ IDNO: 105); an amino acid sequence motif 8 as represented by an amino acidsequence of the formulaIRLD[QKNR]MEFSEVM[VAIL][IVL]HRM[FHYW][IVFL][RK][LMIV][DPE][DE]LD[IVL][AG][HLIV](SEQ ID NO: 106); an amino acid sequence motif 9 as represented by anamino acid sequence of the formula[ALIV]A[REKD][GKR]L[QKENRD][RK]V[LVI][EQDN]IL[ED][GDE]LQA[VAIL]MEVV[AKR][AILV]I[KNRQ][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][MLIV]P[TS]E(SEQ ID NO: 107); an amino acid sequence motif 10 as represented by anamino acid sequence of the formula[TVSIL][FP[LSIVT][TS][WLYFVI]SRVRI[HRK][HYWF][LVI]EMKF[VDILE][GQAN][AG]A(SEQ ID NO: 108); an amino acid sequence motif 11 as represented by anamino acid sequence of the formula[RK]Q[VLI]A[RN]A[LVI]FAV[LFIV][GRK]AI[AG][AST][VIL][AG]L[ATS][FVLI][ALVI]TGGA(SEQ ID NO: 109); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[QLNIV][RK][QRTNKS][STILV]EL[DQEN]N[TAS][IMLV][QEDN][KR][MIVL]DQLSLQMET[QEDN]S[AED][DAE]M[ED]QA[KQRN][AED]DM[ED][AED] (SEQ ID NO: 110); an aminoacid sequence motif 13 as represented by an amino acid sequence of theformula LWP[QRNK]LQY[DHE]MYSNLI[NDQE][RQKN][MLIV][AVIL][QRNK]VA (SEQ IDNO: 111); an amino acid sequence motif 14 as represented by an aminoacid sequence of the formula[PA][DVELI]A[LVI]LTDP[NSQT][IVL][LIV][LSIVT][GC]MQT[ST][MLIV]LIAELV[ELDIV][FVIL][AST][HQN][PNQ]SS (SEQ ID NO: 112); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[ATS][TVSIL][SGT]A[DGE][ADE]TTQIAITF[HYFW][VIL][ST]AIR[RQEKND]I (SEQ IDNO: 113); an amino acid sequence motif 16 as represented by an aminoacid sequence of the formula LEGA[ED]KVKRLYVFADVVEL[PED][SGT]K (SEQ IDNO: 114); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula P[IVL]TD[TS]GE[VIL]Y[IMVL]LLQ[SGT]SR (SEQID NO: 115); an amino acid sequence motif 18 as represented by an aminoacid sequence of the formulaT[VLI][RSKT]LPG[ST][IVL][MST]WILCR[VIL]L[HVIL][LVI]N[GST] (SEQ ID NO:116); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula[AVIL]V[TR][KR]H[VAIL]EWLN[KT]LL[LVI]Q[AVIL]SAAAQ[GST] (SEQ ID NO: 117);an amino acid sequence motif 20 as represented by an amino acid sequenceof the formula [YV][LVI]ALL[FYW]R[ATS]Q[YAWF][LVFI][ILV]K[LGMIV]V[GKR](SEQ ID NO: 118); and an amino acid sequence motif 21 as represented byan amino acid sequence of the formulaAP[GALIV]A[VIML][AST]AA[KGR][GA]AV[ST][AIL]AG (SEQ ID NO: 119); an aminoacid sequence motif 22 as represented by an amino acid sequence of theformula [ED]D[WYF][EQDN]FE[ILV]PVEDF (SEQ ID NO: 120); an amino acidsequence motif 23 as represented by an amino acid sequence of theformula YS[EHD]LYR[DE][LVI]NQ[VIL]S (SEQ ID NO: 121) an amino acidsequence motif 24 as represented by an amino acid sequence of theformula F[AIV][NRQK]TFMRMT (SEQ ID NO: 122).

In some embodiments the PtIP-50 polypeptide comprises at least one aminoacid sequence motif selected from: an amino acid sequence motif 1 havingat least 90% identity to an amino acid sequence ofAWK[ATS][KS]C[KR]NVA[AVIL][LVI]G[RQKN]E[MLIV][CTS]TTAA[YHWF]I[SAT][EQDN]LQY[DEQN]I[QKNR]VQ[AED]MLQ[EDQN]IA[RQKN][KR]QA[DE]RL[SETD][SGAT]I(SEQ ID NO: 99); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of[EQDN][SAT]YDQ[ESDT][LFIV][KR]Q[LFIV][KQRN]LF[IVL][AQN]QN[KERD]ILG[SGT]YLL[EQDN]QN[RK]AFA[EAD][KR]E[RK]DM[ED][VAIL]FHS (SEQ ID NO: 100); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence ofF[HRK]DRN[EKDR][TASGERDNKQ]E[VPIL]L[HERKD]YEA[AG][TVSIL][PG]L[DVELI]Y[HQN]YAY[NRQK]LDTG[EAD]TT[LVI][TS]NLPS(SEQ ID NO: 101); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence ofADL[ST][SNTQ][YFW]TE[MLIV][VAIL][TS][QEDN]MDMRTTR[LMIV]L[LVI][EAD]LIKVL[HNYWF][IMVL]QN[AGVIL]ALMY[QEDN]YLS[EPD][PA] (SEQ ID NO: 102); an amino acidsequence motif 5 having at least 90% identity to an amino acid sequenceof PFT[RNTKQS]WRLR[LVI]SASA[EQDN]EN[EKQDRN]GLAFP (SEQ ID NO: 103); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of DW[LDIVE]IF[VEILD]NE[VIL]E[AG]VA[EAD]QMP[TESD]EVSE (SEQID NO: 104); an amino acid sequence motif 7 having at least 90% identityto an amino acid sequence ofV[TNSQ]M[EDQN]TVW[GRK][MLIV]L[VIL]Q[HQN]E[HQN]AA[IVL][LQIVN]GL[LMIV][RQKN]LG[PA][SAPT][FSWYT]D[FILV][TRVSKIL]RT[YFW]VVKDIPV[DSET]LLL[DHE]G (SEQ IDNO: 105); an amino acid sequence motif 8 having at least 90% identity toan amino acid sequence ofIRLD[QKNR]MEFSEVM[VAIL][IVL]HRM[FHYW][IVFL][RK][LMIV][DPE][DE]LD[IVL][AG][HLIV](SEQ ID NO: 106); an amino acid sequence motif 9 having at least 90%identity to an amino acid sequence of[ALIV]A[REKD][GKR]L[QKENRD][RK]V[LVI][EQDN]IL[ED][GDE]LQA[VAIL]MEVV[AKR][AILV]I[KNRQ][DE]LVESLQE[LIV]GQLV[DE]AP[ED]MP[DE][MLIV]P[TS]E (SEQ ID NO:107); an amino acid sequence motif 10 having at least 90% identity to anamino acid sequence of[TVSIL][FP[LSIVT][TS][WLYFVI]SRVRI[HRK][HYWF][LVI]EMKF[VDILE][GQAN][AG]A(SEQ ID NO: 108); an amino acid sequence motif 11 having at least 90%identity to an amino acid sequence of[RK]Q[VLI]A[RN]A[LVI]FAV[LFIV][GRK]AI[AG][AST][VIL][AG]L[ATS][FVLI][ALVI]TGGA(SEQ ID NO: 109); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of[QLNIV][RK][QRTNKS][STILV]EL[DQEN]N[TAS][IMLV][QEDN][KR][MIVL]DQLSLQMET[QEDN]S[AED][DAE]M[ED]QA[KQRN][AED]DM[ED][AED] (SEQ ID NO: 110); an aminoacid sequence motif 13 having at least 90% identity to an amino acidsequence of LWP[QRNK]LQY[DHE]MYSNLI[NDQE][RQKN][MLIV][AVIL][QRNK]VA (SEQID NO: 111); an amino acid sequence motif 14 having at least 90%identity to an amino acid sequence of[PA][DVELI]A[LVI]LTDP[NSQT][IVL][LIV][LSIVT][GC]MQT[ST][MLIV]LIAELV[ELDIV][FVIL][AST][HQN][PNQ]SS (SEQ ID NO: 112); an amino acid sequence motif 15 havingat least 90% identity to an amino acid sequence of[ATS][TVSIL][SGT]A[DGE][ADE]TTQIAITF[HYFW][VIL][ST]AIR[RQEKND]I (SEQ IDNO: 113); an amino acid sequence motif 16 having at least 90% identityto an amino acid sequence of LEGA[ED]KVKRLYVFADVVEL[PED][SGT]K (SEQ IDNO: 114); an amino acid sequence motif 17 having at least 90% identityto an amino acid sequence of P[IVL]TD[TS]GE[VIL]Y[IMVL]LLQ[SGT]SR (SEQID NO: 115); an amino acid sequence motif 18 having at least 90%identity to an amino acid sequence ofT[VLI][RSKT]LPG[ST][IVL][MST]WILCR[VIL]L[HVIL][LVI]N[GST] (SEQ ID NO:116); an amino acid sequence motif 19 having at least 90% identity to anamino acid sequence of[AVIL]V[TR][KR]H[VAIL]EWLN[KT]LL[LVI]Q[AVIL]SAAAQ[GST] (SEQ ID NO: 117);an amino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of [YV][LVI]ALL[FYW]R[ATS]Q[YAWF][LVFI][ILV]K[LGMIV]V[GKR](SEQ ID NO: 118); and an amino acid sequence motif 21 having at least90% identity to an amino acid sequence ofAP[GALIV]A[VIML][AST]AA[KGR][GA]AV[ST][AIL]AG (SEQ ID NO: 119); an aminoacid sequence motif 22 having at least 90% identity to an amino acidsequence of [ED]D[WYF][EQDN]FE[ILV]PVEDF (SEQ ID NO: 120); an amino acidsequence motif 23 having at least 90% identity to an amino acid sequenceof YS[EHD]LYR[DE][LVI]NQ[VIL]S (SEQ ID NO: 121), and an amino acidsequence motif 24 having at least 90% identity to an amino acid sequenceof F[AIV][NRQK]TFMRMT (SEQ ID NO: 122).

In some embodiments the PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus at least one amino acid sequence motifselected from: motif 23, motif 8, motif 16, motif 18, motif 14, motif19, motif 20, motif 13, motif 2, motif 12, motif 11, motif 21, motif 9,motif 6, motif 1, motif 4, motif 7, motif 22, motif 10, motif 17, motif3, motif 24, motif 5, and motif 15.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having predominantly a nonconservedsecondary structure; a Region B of between about 380 to about 465 aminoacids in length having a consensus secondary structure comprising 8 to10 segments of predominately alpha helical structure; and a Region C ofbetween about 150 to about 180 amino acids in length having a consensussecondary structure comprising 6 to 8 segments of predominately betastrand structure. As used herein “predominantly a nonconserved secondarystructure” means that the regions of secondary structure don'tconsistently align within the family of PtIP-50 polypeptides. As usedherein “predominately alpha helical structure” means that secondarystructure prediction may have one or more gap of between 1 to 4 aminoacids of coil and/or beta strand structure intervening in the alphahelix structure. As used herein “predominately beta strand structure”means that secondary structure prediction may have a gap of between 1 to4 amino acids of coil and/or alpha helix structure intervening in thebeta strand structure. In some embodiments the secondary structure isgenerated by the PSIPRED, top ranked secondary structure predictionmethod (Jones D T. (1999) J. Mol. Biol. 292: 195-202).

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a predominantly nonconservedsecondary structure; a Region B of between about 380 to about 465 aminoacids in length having a consensus secondary structure comprising ninesegments of predominately alpha helical structure; and a Region C ofbetween about 150 to about 180 amino acids in length having a consensussecondary structure comprising seven segments of predominately betastrand structure.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a flexible consensus secondarystructure, wherein the Region A comprises a conserved beta strand 1(β1a) of between about 4 and about 12 amino acids in length within aboutamino acid residue 50 to about amino acid residue 130 from theN-terminus of the PtIP-50 polypeptide; a Region B of between about 380to about 465 amino acids in length having a consensus secondarystructure comprising nine segments of predominately alpha helicalstructure; and a Region C of between about 150 to about 180 amino acidsin length having a consensus secondary structure comprising sevensegments of predominately beta strand structure. As used herein, theterm “about” when used in the context of the lower/upper limit of thelength of a secondary structural element means the greater of −/+aninteger of up to −/+20% of the length of the secondary structuralelement or −/+1 amino acid. By means of example, a secondary structureelement of between about 3 amino acids and about 23 amino acids inlength means a secondary structure element of between 2 and 27 aminoacids in length.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a flexible consensus secondarystructure, wherein the Region A comprises a conserved beta strand 1(β1a) of between about 4 and about 12 amino acids in length, a coil ofbetween about 3 and about 18 amino acids in length and a beta strand 2(β1b) of between about 4 and about 32 amino acids in length, withinabout amino acid residue 50 to about amino acid residue 165 from theN-terminus of the PtIP-50 polypeptide; a Region B of between about 380to about 465 amino acids in length having a consensus secondarystructure comprising nine segments of predominately alpha helicalstructure; and a Region C of between about 150 to about 180 amino acidsin length having a consensus secondary structure comprising sevensegments of predominately beta strand structure.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a predominantly nonconservedsecondary structure; a Region B of between about 380 to about 465 aminoacids in length having a consensus secondary structure sequentiallycomprising i) an alpha helix-1 of between about 10 and about 26 aminoacids in length; ii) a coil-1 of between about 2 and about 8 amino acidsin length flanked by alpha helix-1 and alpha helix-2; iii) an alphahelix-2 of between about 15 and about 24 amino acids in length; iv) acoil-2 of between about 4 and about 14 amino acids in length flanked byalpha helix-2 and alpha helix-3; v) an alpha helix 3 of between about 10and about 17 amino acids in length; vi) a coil-3 of between about 11 andabout 17 amino acids in length flanked by alpha helix-3 and alphahelix-4; vii) an alpha helix-4 of between about 24 and about 30 aminoacids in length; viii) a coil-4 of between about 4 and about 13 aminoacids in length flanked by alpha helix-4 and alpha helix-5; ix) an alphahelix-5 of between about 48 and about 58 amino acids in length; x) acoil-5 of between about 10 and about 15 amino acids in length flanked byalpha helix-5 and alpha helix-6; xi) an alpha helix-6 of between about13 and about 15 amino acids in length; xii) a coil-6 of between about 3and about 10 amino acids in length flanked by alpha helix-6 and alphahelix-7; xiii) an alpha helix-7 of between about 49 and about 59 aminoacids in length; xiv) a coil-7 of between about 0 and about 9 aminoacids in length flanked by alpha helix-7 and alpha helix-8; xv) an alphahelix-8 of between about 29 and about 36 amino acids in length; xvi) acoil-8 of between about 11 and about 16 amino acids in length flanked byalpha helix-8 and alpha helix-9; xvii) an alpha helix-9 of between about16 and about 22 amino acids in length; xviii) a coil-9 of between about5 and about 9 amino acids in length flanked by alpha helix-9 and RegionC; and a Region C of between about 150 to about 180 amino acids inlength having a consensus secondary structure comprising seven segmentsof predominately beta strand structure.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a predominantly nonconservedsecondary structure; a Region B of between about 380 to about 465 aminoacids in length having a consensus secondary structure comprising ninesegments of predominately alpha helical structure; and a Region C ofbetween about 150 to about 180 amino acids in length having a consensussecondary structure sequentially comprising i) a beta strand-1 (β1) ofbetween about 5 amino acids and about 7 amino acids in length; ii) acoil of between about 12 amino acids and about 17 amino acids in length;iii) a beta strand-2 (β2) of between about 11 amino acids and about 15amino acids in length; iv) a coil of between about 7 amino acids andabout 23 amino acids in length; v) a beta strand-3 (β3) of between about5 amino acids and about 8 amino acids in length; vi) a coil of betweenabout 13 amino acids and about 23 amino acids in length; vii) a betastrand-4 (β4) of between about 3 amino acids and about 7 amino acids inlength; viii) a coil of between about 3 amino acids and about 8 aminoacids in length; ix) a beta strand-5 (β5) of between about 4 amino acidsand about 10 amino acids in length; x) a coil of between about 24 aminoacids and about 28 amino acids in length; xi) a beta strand-6 (β6) ofbetween about 5 amino acids and about 7 amino acids in length; xii) acoil of between about 17 amino acids and about 23 amino acids in length;and xiii) a beta strand-1 (β7) of between about 5 amino acids and about7 amino acids in length.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a predominantly nonconservedsecondary structure; a Region B of between about 380 to about 465 aminoacids in length having a consensus secondary structure sequentiallycomprising i) an alpha helix-1 of between about 10 and about 26 aminoacids in length; ii) a coil-1 of between about 2 and about 8 amino acidsin length flanked by alpha helix-1 and alpha helix-2; iii) an alphahelix-2 of between about 15 and about 24 amino acids in length; iv) acoil-2 of between about 4 and about 14 amino acids in length flanked byalpha helix-2 and alpha helix-3; v) an alpha helix 3 of between about 10and about 17 amino acids in length; vi) a coil-3 of between about 11 andabout 17 amino acids in length flanked by alpha helix-3 and alphahelix-4; vii) an alpha helix-4 of between about 24 and about 30 aminoacids in length; viii) a coil-4 of between about 4 and about 13 aminoacids in length flanked by alpha helix-4 and alpha helix-5; ix) an alphahelix-5 of between about 48 and about 58 amino acids in length; x) acoil-5 of between about 10 and about 15 amino acids in length flanked byalpha helix-5 and alpha helix-6; xi) an alpha helix-6 of between about13 and about 15 amino acids in length; xii) a coil-6 of between about 3and about 10 amino acids in length flanked by alpha helix-6 and alphahelix-7; xiii) an alpha helix-7 of between about 49 and about 59 aminoacids in length; xiv) a coil-7 of between about 0 and about 9 aminoacids in length flanked by alpha helix-7 and alpha helix-8; xv) an alphahelix-8 of between about 29 and about 36 amino acids in length; xvi) acoil-8 of between about 11 and about 16 amino acids in length flanked byalpha helix-8 and alpha helix-9; xvii) an alpha helix-9 of between about16 and about 22 amino acids in length; xviii) a coil-9 of between about5 and about 9 amino acids in length flanked by alpha helix-9 and betastrand-1 (β1) of Region C; and a Region C of between about 150 to about180 amino acids in length having a consensus secondary structuresequentially comprising i) a beta strand-1 (β1) of between about 5 aminoacids and about 7 amino acids in length; ii) a coil of between about 12amino acids and about 17 amino acids in length; iii) a beta strand-2(β2) of between about 11 amino acids and about 15 amino acids in length;iv) a coil of between about 7 amino acids and about 23 amino acids inlength; v) a beta strand-3 (β3) of between about 5 amino acids and about8 amino acids in length; vi) a coil of between about 13 amino acids andabout 23 amino acids in length; vii) a beta strand-4 (β4) of betweenabout 3 amino acids and about 7 amino acids in length; viii) a coil ofbetween about 3 amino acids and about 8 amino acids in length; ix) abeta strand-5 (β5) of between about 4 amino acids and about 10 aminoacids in length; x) a coil of between about 24 amino acids and about 28amino acids in length; xi) a beta strand-6 (β6) of between about 5 aminoacids and about 7 amino acids in length; xii) a coil of between about 17amino acids and about 23 amino acids in length; and xiii) a betastrand-1 (β7) of between about 5 amino acids and about 7 amino acids inlength.

In some embodiments a PtIP-50 polypeptide comprises sequentially fromthe N-terminus to the C-terminus: a Region A of between about 200 toabout 300 amino acids in length having a flexible consensus secondarystructure, wherein the Region A comprises a conserved beta strand 1(β1a) of between about 4 and about 12 amino acids in length within aboutamino acid residue 50 to about amino acid residue 130 from theN-terminus of the PtIP-50 polypeptide; a Region B of between about 380to about 465 amino acids in length having a consensus secondarystructure sequentially comprising i) an alpha helix-1 of between about10 and about 26 amino acids in length; ii) a coil-1 of between about 2and about 8 amino acids in length flanked by alpha helix-1 and alphahelix-2; iii) an alpha helix-2 of between about 15 and about 24 aminoacids in length; iv) a coil-2 of between about 4 and about 14 aminoacids in length flanked by alpha helix-2 and alpha helix-3; v) an alphahelix 3 of between about 10 and about 17 amino acids in length; vi) acoil-3 of between about 11 and about 17 amino acids in length flanked byalpha helix-3 and alpha helix-4; vii) an alpha helix-4 of between about24 and about 30 amino acids in length; viii) a coil-4 of between about 4and about 13 amino acids in length flanked by alpha helix-4 and alphahelix-5; ix) an alpha helix-5 of between about 48 and about 58 aminoacids in length; x) a coil-5 of between about 10 and about 15 aminoacids in length flanked by alpha helix-5 and alpha helix-6; xi) an alphahelix-6 of between about 13 and about 15 amino acids in length; xii) acoil-6 of between about 3 and about 10 amino acids in length flanked byalpha helix-6 and alpha helix-7; xiii) an alpha helix-7 of between about49 and about 59 amino acids in length; xiv) a coil-7 of between about 0and about 9 amino acids in length flanked by alpha helix-7 and alphahelix-8; xv) an alpha helix-8 of between about 29 and about 36 aminoacids in length; xvi) a coil-8 of between about 11 and about 16 aminoacids in length flanked by alpha helix-8 and alpha helix-9; xvii) analpha helix-9 of between about 16 and about 22 amino acids in length;xviii) a coil-9 of between about 5 and about 9 amino acids in lengthflanked by alpha helix-9 and beta strand-1 (β1) of Region C; and aRegion C of between about 150 to about 180 amino acids in length havinga consensus secondary structure sequentially comprising i) a betastrand-1 (β1) of between about 5 amino acids and about 7 amino acids inlength; ii) a coil of between about 12 amino acids and about 17 aminoacids in length; iii) a beta strand-2 (β2) of between about 11 aminoacids and about 15 amino acids in length; iv) a coil of between about 7amino acids and about 23 amino acids in length; v) a beta strand-3 (β3)of between about 5 amino acids and about 8 amino acids in length; vi) acoil of between about 13 amino acids and about 23 amino acids in length;vii) a beta strand-4 (β4) of between about 3 amino acids and about 7amino acids in length; viii) a coil of between about 3 amino acids andabout 8 amino acids in length; ix) a beta strand-5 (β5) of between about4 amino acids and about 10 amino acids in length; x) a coil of betweenabout 24 amino acids and about 28 amino acids in length; xi) a betastrand-6 (β6) of between about 5 amino acids and about 7 amino acids inlength; xii) a coil of between about 17 amino acids and about 23 aminoacids in length; and xiii) a beta strand-1 (β7) of between about 5 aminoacids and about 7 amino acids in length.

In some embodiments a PtIP-50 polypeptide has a calculated molecularweight of between about 70kD and about 120kD, between about 75kD andabout 110kD, between about 80kD and about 105kD, or between about 85kDand about 105kD.

In some embodiments the PtIP-50 polypeptide has a modified physicalproperty. As used herein, the term “physical property” refers to anyparameter suitable for describing the physical-chemical characteristicsof a protein. As used herein, “physical property of interest” and“property of interest” are used interchangeably to refer to physicalproperties of proteins that are being investigated and/or modified.Examples of physical properties include, but are not limited to netsurface charge and charge distribution on the protein surface, nethydrophobicity and hydrophobic residue distribution on the proteinsurface, surface charge density, surface hydrophobicity density, totalcount of surface ionizable groups, surface tension, protein size and itsdistribution in solution, melting temperature, heat capacity, and secondvirial coefficient. Examples of physical properties also include, butare not limited to solubility, folding, stability, and digestibility. Insome embodiments the PtIP-50 polypeptide has increased digestibility ofproteolytic fragments in an insect gut. Models for digestion bysimulated gastric fluids are known to one skilled in the art (Fuchs, R.L. and J. D. Astwood. Food Technology 50: 83-88, 1996; Astwood, J. D.,et al Nature Biotechnology 14: 1269-1273, 1996; Fu T J et al J. AgricFood Chem. 50: 7154-7160, 2002).

PtIP-65 Polypeptides

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ IDNO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42,wherein the PtIP-65 polypeptide, in combination with a PtIP-50polypeptide, has insecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24,SEQ ID NO: 26 or SEQ ID NO: 29, wherein the PtIP-65 polypeptide, incombination with a PtIP-50 polypeptide, has insecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 27,SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO:36, wherein the PtIP-65 polypeptide in combination with a PtIP-50polypeptide has insecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 28,SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33, wherein the PtIP-65polypeptide, in combination with a PtIP-50 polypeptide, has insecticidalactivity. In some embodiments the PtIP-65 polypeptide comprises an aminoacid sequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identity to the amino acid sequence of SEQ ID NO: 41 or SEQID NO: 42, wherein the PtIP-65 polypeptide, in combination with aPtIP-50 polypeptide, has insecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 38 or SEQ ID NO:40, wherein the PtIP-65 polypeptide, in combination with a PtIP-50polypeptide, has insecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 37, wherein thePtIP-65 polypeptide, in combination with a PtIP-50 polypeptide, hasinsecticidal activity.

In some embodiments the PtIP-65 polypeptide comprises an amino acidsequence having at least 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%,57%, 58%, 59%, 60%, 81%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identity to the amino acid sequence of SEQ ID NO: 39, wherein thePtIP-65 polypeptide, in combination with a PtIP-50 polypeptide, hasinsecticidal activity.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Division Pteridophyta. The phylogeny of ferns as usedherein is based on the classification for extant ferns by A. R. Smith etal, TAXON, 55:705-731 (2006). The consensus phylogeny based on theclassification by A. R. Smith is shown in FIG. 1. Additional informationon the phylogeny of ferns can be found atmobot.org/MOBOT/research/APweb/(which can be accessed using the “www”prefix) and Schuettpelz E. and Pryer K. M., TAXON 56: 1037-1050 (2007)based on three plastid genes. Additional fern and other primitive plantspecies can be found at homepages.caverock.net.nz/˜byfern/list.htm(which can be accessed using the http:// prefix).

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Class Psilotopsida. In some embodiments the PtIP-65polypeptide is derived from a fern species in the Class Psilotopsida,Order Psilotales. In some embodiments the PtIP-65 polypeptide is derivedfrom a fern species in the Class Psilotopsida, Order Ophioglossales. Insome embodiments the PtIP-65 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales, Family Psilotaceae. Insome embodiments the PtIP-65 polypeptide is derived from a fern speciesin the Class Psilotopsida, Order Ophioglossales Family Ophioglossaceae.In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Genus Ophioglossum L., Botrychium, Botrypus,Helminthostachys, Ophioderma, Cheiroglossa, Sceptridium or Mankyua. Insome embodiments the PtIP-65 polypeptide is derived from a fern speciesin the Ophioglossum L. Genus is selected from but not limited toOphioglossum californicum, Ophioglossum coriaceum, Ophioglossumcostatum, Ophioglossum crotalophoroides, Ophioglossum engelmannii,Ophioglossum falcatum, Ophioglossum gomezianum, Ophioglossum gramineum,Ophioglossum kawarnurae, Ophioglossum lusitanicum, Ophioglossumnamegatae, Ophioglossum nudicaule, Ophioglossum palmatum, Ophioglossumparvum, Ophioglossum pedunculosum, Ophioglossum pendulum, Ophioglossumpetiolatum, Ophioglossum pusillum, Ophioglossum reticulatum,Ophioglossum richardsiae, Ophioglossum thermale, and Ophioglossumvulgatum.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Class Polypodiopsida/Pteridopsida. In some embodiments the PtIP-65polypeptide is derived from a fern species in the Order Osmundales(royal ferns); Family Osmundaceae. In some embodiments the PtIP-65polypeptide is derived from a fern species in the Order Hymenophyllales(filmy ferns and bristle ferns); Family Hymenophyllaceae. In someembodiments the PtIP-65 polypeptide is derived from a fern species inthe Order Gleicheniales; Family Gleicheniaceae, Family Dipteridaceael orFamily Matoniaceae. In some embodiments the PtIP-65 polypeptide isderived from a fern species in the Order Schizaeales; FamilyLygodiaceae, Family Anemiaceae or Family Schizaeaceae. In someembodiments the PtIP-65 polypeptide is derived from a fern species inthe Order Salviniales; Family Marsileaceae or Family Salviniaceae. Insome embodiments the PtIP-65 polypeptide is derived from a fern speciesin the Order Cyatheales; Family Thyrsopteridaceae, Family Loxsomataceae,Family Culcitaceae, Family Plagiogyriaceae, Family Cibotiaceae, FamilyCyatheaceae, Family Dicksoniaceae or Family Metaxyaceae.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales; Family Lindsaeaceae, FamilySaccolomataceae, Family Cystodiaceae, Family Dennstaedtiaceae, FamilyPteridaceae, Family Aspleniaceae, Family Thelypteridaceae, FamilyWoodsiaceae, Family Onocleaceae, Family Blechnaceae, FamilyDryopteridaceae, Family Lomariopsidaceae, Family Tectariaceae, FamilyOleandraceae, Family Davalliaceae or Family Polypodiaceae.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Pteridaceae, Genus Adiantaceaeselected from but not limited to Adiantum aethiopicum, Adiantumaleuticum, Adiantum bonatianum, Adiantum cajennense, Adiantumcapillus-junonis, Adiantum capillus-veneris, Adiantum caudatum, Adiantumchienfi, Adiantum chilense, Adiantum cuneatum, Adiantum cunninghamfi,Adiantum davidii, Adiantum diaphanum, Adiantum edentulum, Adiantumedgeworthfi, Adiantum excisum, Adiantum fengianum, Adiantum fimbriatum,Adiantum flabellulaturn, Adiantum formosanum, Adiantum formosum,Adiantum fulvum, Adiantum gravesfi, Adiantum hispidulum, Adiantuminduratum, Adiantum jordanfi, Adiantum juxtapositum, Adiantumlatifolium, Adiantum leveillei, Adiantum lianxianense, Adiantummalesianum, Adiantum mariesfi, Adiantum monochlamys, Adiantummyriosorum, Adiantum obliquum, Adiantum ogasawarense, Adiantum pedatum,Adiantum pentadactylon, Adiantum peruvianum, Adiantum philippense,Adiantum princeps, Adiantum pubescens, Adiantum raddianum, Adiantumreniforme, Adiantum roborowskii, Adiantum serratodentatum, Adiantumsinicum, Adiantum soboliferum, Adiantum subcordatum, Adiantum tenerum,Adiantum terminatum, Adiantum tetraphyllum, Adiantum venustum, Adiantumviridescens, and Adiantum viridimontanurn.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL. In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Aspleniaceae, Genus AspleniumL selected from but not limited to Asplenium abbreviatum, Aspleniumabrotanoides, Asplenium abscissum var. subaequilaterale, Aspleniumabscissum, Asplenium achilleifolium, Asplenium acuminatum, Aspleniumadiantifrons, Asplenium adiantoides, Asplenium adiantoides var.squamulosum, Asplenium adiantum-nigrum L., Asplenium adiantum-nigrumvar. adiantum-nigrum, Asplenium adiantum-nigrum var. yuanurn, Aspleniumadnatum, Asplenium aethiopicum, Asplenium affine, Asplenium affine var.affine, Asplenium affine var. gilpinae, Asplenium affine var. mettenii,Asplenium affine var. pecten, Asplenium africanum, Asplenium afzelii,Asplenium aitchisonii, Asplenium alatulum, Asplenium alatum, Aspleniumalfredii, Asplenium altajense, Asplenium amabile, Aspleniumambohitantelense, Asplenium anceps var. proliferum, Asplenium andapense,Asplenium andersonii, Asplenium angustatum, Asplenium angustum,Asplenium anisophyllum, Asplenium annetii, Asplenium antiquum, Aspleniumantrophyoides, Asplenium apertura, Asplenium apogamum, Aspleniumaquaticum, Asplenium arboreum, Asplenium arcanum, Asplenium arcuatum,Asplenium argentinurn, Asplenium argutum, Asplenium aspidiiforme,Asplenium aspidioides, Asplenium asterolepis, Asplenium auriculariumvar. acutidens, Asplenium auricularium var. subintegerrimum, Aspleniumauriculaturn, Asplenium auriculaturn var. aequilaterale, Aspleniumauritum fo. diversifolium, Asplenium auritum fo. diversifolium,Asplenium auritum fo. nana, Asplenium auritum, Asplenium auritum var.auriculaturn, Asplenium auritum var. auritum, Asplenium auritum var.bipinnatifidum, Asplenium auritum var. bipinnatisectum, Aspleniumauritum var. davallioides, Asplenium auritum var. macilentum, Aspleniumauritum var. rigidum, Asplenium auritum var. subsimplex, Aspleniumaustrochinense, Asplenium ayopayense, Asplenium badinii, Aspleniumbalense, Asplenium ballivianii, Asplenium bangii, Asplenium bangii,Asplenium barbaense, Asplenium barclayanum, Asplenium barkamense,Asplenium barteri, Asplenium basiscopicum, Asplenium bicrenaturn,Asplenium bifrons, Asplenium bipartitum, Asplenium blastophorum,Asplenium blepharodes, Asplenium blepharophorum, Asplenium boiteaui,Asplenium bolivianum, Asplenium boltonii, Asplenium borealichinense,Asplenium bradei, Asplenium bradeorum, Asplenium bradleyi, Aspleniumbrausei, Asplenium breedlovei, Asplenium buettneri, Asplenium buettnerivar. hildebrandtii, Asplenium bulbiferum, Asplenium bullaturn var.bullaturn, Asplenium bullatum var. shikokianum, Asplenium bullaturn,Asplenium cancellatum, Asplenium capillipes, Asplenium cardiophyllum(Hance), Asplenium caripense, Asplenium carvalhoanum, Aspleniumcastaneoviride, Asplenium castaneum, Asplenium caudatum, Aspleniumceltidifolium (Kunze), Asplenium ceratolepis, Asplenium changputungense,Asplenium chaseanum, Asplenium cheilosorum, Asplenium chengkouense,Asplenium chihuahuense, Asplenium chimantae, Asplenium chimborazense,Asplenium chingianum, Asplenium chlorophyllum, Asplenium chondrophyllum,Asplenium cicutarium, Asplenium cicutarium var. paleaceum, Aspleniumcirrhaturn, Asplenium cladolepton, Asplenium claussenii, Aspleniumcoenobiale, Asplenium commutatum, Asplenium congestum, Aspleniumconquisitum, Asplenium consimile, Asplenium contiguum, Aspleniumcontiguum var. hirtulum, Asplenium corderoi, Asplenium cordovense,Asplenium coriaceum, Asplenium corfifolium, Asplenium correardii,Asplenium costale, Asplenium costale var. robustum, Asplenium cowanii,Asplenium crenulatoserrulatum, Asplenium crenulatum, Aspleniumcrinicaule, Asplenium crinulosum, Asplenium cristatum, Aspleniumctyptolepis Fernald, Asplenium cultrifolium L., Asplenium cuneatiforme,Asplenium cuneatum, Asplenium curvatura, Asplenium cuspidatum, Aspleniumcuspidatum var cuspidatum, Asplenium cuspidatum var. foeniculaceum,Asplenium cuspidatum var. triculum, Asplenium cuspidatum var.tripinnaturn, Asplenium dalhousiae, Asplenium dareoides, Aspleniumdavallioides, Asplenium davisii, Asplenium debile, Asplenium debile,Asplenium decussatum, Asplenium delavayi, Asplenium delicatulum,Asplenium delicatulum var. cocosensis, Asplenium delitescens, Aspleniumdelitescens×laetum, Asplenium densum, Asplenium dentatum L., Aspleniumdentatum L., Asplenium depauperatum, Asplenium deqenense, Aspleniumdianae, Asplenium difforme, Asplenium dilatatum, Asplenium dimidiatum,Asplenium dimidiaturn var. boliviense, Asplenium diplazisorum, Aspleniumdissectum, Asplenium distans, Asplenium divaricatum, Aspleniumdivergens, Asplenium divisissimurn, Asplenium doederleinii, Aspleniumdonnell-smithii, Asplenium dregeanum, Asplenium dulongjiangense,Asplenium duplicatoserratum, Asplenium eatonii, Asplenium ebeneum,Asplenium ebenoides, Asplenium ecuadorense, Asplenium eggersii,Asplenium emarginaturn, Asplenium enaturn, Asplenium ensiforme fo.bicuspe, Asplenium ensiforme fo. ensiforme, Asplenium ensiforme fo.stenophyllum, Asplenium ensiforme, Asplenium erectum var. erectum,Asplenium erectum var. gracile, Asplenium erectum var. usambarense,Asplenium erectum var. zeyheri, &, Asplenium erosum L., Aspleniumescaleroense, Asplenium esculenturn, Asplenium eutecnurn, Aspleniumexcelsum, Asplenium excisum, Asplenium exiguum, Asplenium extensum,Asplenium falcatum, Asplenium falcinellum, Asplenium faurei, Aspleniumfeei, Asplenium fengyangshanense, Asplenium ferulaceum, Aspleniumfibrillosum, Asplenium filix-femina, Asplenium finckii, Aspleniumfinlaysonianurn, Asplenium flabellulatum, Asplenium flabellulatum varflabellulatum, Asplenium flabellulatum var. partitum, Aspleniumflaccidum, Asplenium flavescens, Asplenium flavidum, Aspleniumflexuosum, Asplenium fluminense, Asplenium foeniculaceum, Aspleniumformosanum, Asplenium formosum var. carolinurn, Asplenium formosum var.incultum, Asplenium formosum, Asplenium fournieri, Asplenium fragile,Asplenium fragile var. lomense, Asplenium fragrans, Asplenium fragransvar. foeniculaceum, Asplenium franconis var. gracile, Aspleniumfraxinifolium, Asplenium friesiorum, Asplenium friesiorum var.nesophilum, Asplenium fugax, Asplenium fujianense, Asplenium furcatum,Asplenium furfuraceum, Asplenium fuscipes, Asplenium fuscopubescens,Asplenium galeottii, Asplenium gautieri, Asplenium gemmiferum, Aspleniumgentryi, Asplenium geppii, Asplenium ghiesbreghtii, Asplenium gilliesii,Asplenium gilpinae, Asplenium glanduliserratum, Asplenium glenniei,Asplenium goldmannii, Asplenium gomezianum, Asplenium grande, Aspleniumgrandifolium, Asplenium grandifrons, Asplenium gregoriae, Aspleniumgriffithianum, Asplenium gulingense, Asplenium hainanense, Aspleniumhallbergii, Asplenium hallei, Asplenium hallii, Asplenium hangzhouense,Asplenium haplophyllum, Asplenium harpeodes, Asplenium harpeodes var.glaucovirens, Asplenium harpeodes var. incisura, Asplenium harrisiiJenman, Asplenium harrisonii, Asplenium hastatum, Asplenium hebeiense,Asplenium hemionitideum, Asplenium hemitomurn, Asplenium henryi,Asplenium herpetopteris, Asplenium herpetopteris var herpetopteris,Asplenium herpetopteris var. acutipinnata, Asplenium herpetopteris var.masoulae, Asplenium herpetopteris var. villosum, Asplenium hesperium,Asplenium heterochroum, Asplenium hians, Asplenium hians var.pallescens, Asplenium hoffmannii, Asplenium holophlebium, Aspleniumhondoense, Asplenium horridum, Asplenium hostmannii, Aspleniumhumistratum, Asplenium hypomelas, Asplenium inaequilaterale, Aspleniumincisura, Asplenium incurvaturn, Asplenium indicum, Asplenium indicumvar. indicum, Asplenium indicum var. yoshingagae, Asplenium induratum,Asplenium indusiatum, Asplenium inexpectatum, Asplenium insigne,Asplenium insiticium, Asplenium insolitum, Asplenium integerrimurn,Asplenium interjectum, Asplenium jamesonii, Asplenium jaundeense,Asplenium juglandifolium, Asplenium kangdingense, Asplenium kansuense,Asplenium kassneri, Asplenium kaulfussii, Asplenium kellermanii,Asplenium kentuckiense, Asplenium khullarii, Asplenium kiangsuense,Asplenium kunzeanurn, Asplenium lacerum, Asplenium laciniatum, Aspleniumlaciniatum var. acutipinna, Asplenium laciniatum var. laciniatum,Asplenium laetum fo. minor, Asplenium laetum, Asplenium laetum var.incisoserratum, Asplenium lamprocaulon, Asplenium laserpitiifolium var.morrisonense, Asplenium lastii, Asplenium latedens, Aspleniumlatifolium, Asplenium laui, Asplenium laurentii, Asplenium leandrianurn,Asplenium lechleri, Asplenium leiboense, Asplenium lepidorachis,Asplenium leptochlamys, Asplenium leptophyllum, Asplenium levyi,Asplenium lindbergii, Asplenium lindeni, Asplenium lineatum, Aspleniumlividum, Asplenium lobatum, Asplenium lobulaturn, Asplenium lokohoense,Asplenium longicauda, Asplenium longicaudaturn, Asplenium longifolium,Asplenium longisorum, Asplenium longjinense, Asplenium lorentzii,Asplenium loriceum, Asplenium loxogrammoides, Asplenium lugubre,Asplenium lunulatum, Asplenium lunulatum var. pteropus, Aspleniumlushanense, Asplenium lydgatei, Asplenium macilentum, Asplenium macraei,Asplenium macrodictyon, Asplenium macrophlebium, Asplenium macrophyllum,Asplenium macropterum, Asplenium macrosorum, Asplenium macrotis,Asplenium macrurum, Asplenium mainlingense, Asplenium mangindranense,Asplenium mannii, Asplenium marginatum L., Asplenium marojejyense,Asplenium martianum, Asplenium matsumurae, Asplenium mauritiensisLorence, Asplenium maximum, Asplenium, ii, Asplenium megalura, Aspleniummegaphyllum, Asplenium meiotomurn, Asplenium melanopus, Aspleniummembranifolium, Asplenium meniscioides, Asplenium mesosorum, Aspleniummexicanurn, Asplenium micropaleatum, Asplenium microtum, Aspleniummildbraedii, Asplenium mildei, Asplenium minimum, Asplenium minutum,Asplenium miradorense, Asplenium miyunense, Asplenium moccenianurn,Asplenium mocquetysii, Asplenium modestum, Asplenium monanthemum var.menziesii, Asplenium monanthes L., Asplenium monanthes var monanthes,Asplenium monanthes var. castaneum, Asplenium monanthes var. wagneri,Asplenium monanthes var. yungense, Asplenium monodon, Aspleniummontanum, Asplenium mosetenense, Asplenium moupinense, Aspleniummucronatum, Asplenium munchii, Asplenium muticum, Asplenium myapteron,Asplenium myriophyllu, Asplenium nakanoanum, Asplenium nanchuanense,Asplenium nemorale, Asplenium neolaserpitiifolium, Aspleniumneomutijugum, Asplenium neovarians, Asplenium nesii, Aspleniumnesioticum, Asplenium nidus L., Asplenium nigricans, Aspleniumniponicum, Asplenium normale, Asplenium normale var. angustum, Aspleniumobesum, Asplenium oblongatum, Asplenium oblongifolium, Aspleniumobovatum, Asplenium obscurum, Asplenium obscurum var. angustum,Asplenium obtusatum var. obtusatum, Asplenium obtusatum var. sphenoides,Asplenium obtusifolium L., Asplenium obtusissimum, Asplenium obversum,Asplenium ochraceum, Asplenium oellgaardii, Asplenium ofeliae, Aspleniumoldhami, Asplenium oligosorum, Asplenium olivaceum, Asplenium onopterisL., Asplenium onustum, Asplenium ortegae, Asplenium otites, Aspleniumpalaciosii, Asplenium palmeri, Asplenium partitum, Asplenium parvisorum,Asplenium parviusculum, Asplenium parvulum, Asplenium patens, Aspleniumpaucifolium, Asplenium paucijugum, Asplenium paucivenosum, Aspleniumpearcei, Asplenium pekinense, Asplenium pellucidum, Asplenium pendulum,Asplenium petiolulatum, Asplenium phyllitidis, Aspleniumpimpinellifolium, Asplenium pinnatifidum, Asplenium pinnatum, Aspleniumplatyneuron, Asplenium platyneuron var. bacculum-rubrum, Aspleniumplatyneuron var. incisum, Asplenium platyphyllum, Asplenium plumbeum,Asplenium poloense, Asplenium polymeris, Asplenium polymorphum,Asplenium polyodon, Asplenium polyodon var. knudsenii, Aspleniumpolyodon var. nitidulum, Asplenium polyodon var. sectum, Aspleniumpolyodon var. subcaudatum, Asplenium polyphyllum, Asplenium poolii,Asplenium poolii fo. simplex, Asplenium poolii var. linearipinnatum,Asplenium potosinum, Asplenium potosinum var. incisum, Aspleniumpraegracile, Asplenium praemorsum, Asplenium preussii, Aspleniumpringleanum, Asplenium pringlei, Asplenium prionitis, Aspleniumprocerum, Asplenium progrediens, Asplenium projectum, Aspleniumprolongatum, Asplenium propinquum, Asplenium protensum, Aspleniumpseudoangustum, Asplenium pseudoerectum, Asplenium pseudofontanum,Asplenium pseudolaserpitiifolium, Asplenium pseudonormale, Aspleniumpseudo pellucidum, Asplenium pseudopraemorsum, Asplenium pseudovarians,Asplenium pseudowilfordii, Asplenium pseudowrightii, Aspleniumpsilacrum, Asplenium pteropus, Asplenium pubirhizoma, Aspleniumpulchellum, Asplenium pulchellum var. subhorizontale, Aspleniumpulcherrimum, Asplenium pulicosum, Asplenium pulicosum var. maius,Asplenium pululahuae, Asplenium pumilum, Asplenium pumilum var.hymenophylloides, Asplenium pumilum var. laciniatum, Aspleniumpurdieanum, Asplenium purpurascens, Asplenium pyramidatum, Aspleniumqiujiangense, Asplenium quercicola, Asplenium quitense, Aspleniumraddianum, Asplenium radiatum, Asplenium radicans L., Aspleniumradicans, Asplenium radicans var. costaricense, Asplenium radicans var.partitum, Asplenium radicans var. radicans, Asplenium radicans var.uniseriale, Asplenium recumbens, Asplenium reflexum, Asplenium regularevar. latior, Asplenium repandulum, Asplenium repens, Asplenium repente,Asplenium resiliens, Asplenium retusulum, Asplenium rhipidoneuron,Asplenium rhizophorum L., Asplenium rhizophyllum, Asplenium rhizophyllumL., Asplenium rhizophyllum var. proliferum, Asplenium rhomboideum,Asplenium rigidum, Asplenium riparium, Asplenium rivale, Aspleniumrockii, Asplenium roemerianum, Asplenium roemerianum var. mindensis,Asplenium rosenstockianum, Asplenium rubinum, Asplenium ruizianum,Asplenium rusbyanum, Asplenium ruta-muraria L., Asplenium ruta-murariavar. cryptolepis, Asplenium rutaceum, Asplenium rutaceum var.disculiferum, Asplenium rutaefolium, Asplenium rutifolium, Aspleniumsalicifolium L., Asplenium salicifolium var. aequilaterale, Aspleniumsalicifolium var. salicifolium, Asplenium sampsoni, Asplenium sanchezii,Asplenium sanderi, Asplenium sandersonii, Asplenium sanguinolentum,Asplenium sarelii, Asplenium sarelii var. magnum, Asplenium sarelii var.sarelii, Asplenium saxicola, Asplenium scalifolium, Aspleniumscandicinum, Asplenium schizophyllum, Asplenium schkuhrii, Aspleniumsciadophilum, Asplenium scolopendrium L., Asplenium scortechinii,Asplenium seileri, Asplenium semipinnatum, Asplenium septentrionale,Asplenium serra, Asplenium serra var. imrayanum, Aspleniumserratissimum, Asplenium serratum L., Asplenium serratum var. caudatum,Asplenium serricula, Asplenium sessilifolium, Asplenium sessilifoliumvar. guatemalense, Asplenium sessilifolium var. minus, Aspleniumsessilifolium var. occidentale, Asplenium sessilipinnum, Aspleniumsetosum, Asplenium shepherdii, Asplenium shepherdii var. bipinnatum,Asplenium shepherdii var. flagelliferum, Asplenium shikokianum,Asplenium simii, Asplenium simonsianum, Asplenium sintenisii, Aspleniumskinneri, Asplenium skinneri, Asplenium sodiroi, Aspleniumsoleirolioides, Asplenium solidum var. stenophyllum, Asplenium solmsii,Asplenium sp.-N.-Halle-2234, Asplenium spathulinum, Aspleniumspectabile, Asplenium speluncae, Asplenium sphaerosporum, Aspleniumsphenotomum, Asplenium spinescens, Asplenium splendens, Aspleniumsprucei, Asplenium squamosum L., Asplenium standleyi, Aspleniumstellatum, Asplenium stenocarpum, Asplenium stoloniferum, Aspleniumstolonipes, Asplenium striatum L., Asplenium stuebelianum, Aspleniumstuhlmannii, Asplenium suave, Asplenium subalatum, Aspleniumsubcrenatum, Asplenium subdigitatum, Asplenium subdimidiatum, Aspleniumsubintegrum, Asplenium sublaserpitiifolium, Asplenium sublongum,Asplenium subnudum, Asplenium suborbiculare, Asplenium subtenuifolium,Asplenium subtile, Asplenium subtoramanum, Asplenium subtrapezoideum,Asplenium subvarians, Asplenium sulcatum, Asplenium sylvaticum,Asplenium szechuanense, Asplenium taiwanense, Asplenium tenerrimum,Asplenium tenerum, Asplenium tenuicaule, Asplenium tenuifolium,Asplenium tenuifolium var. minor, Asplenium tenuifolium var.tenuifolium, Asplenium tenuissimum, Asplenium ternatum, Aspleniumtheciferum, Asplenium theciferum var. concinnum, Asplenium thunbergii,Asplenium tianmushanense, Asplenium tianshanense, Asplenium tibeticum,Asplenium tocoraniense, Asplenium toramanum, Asplenium trapezoideum,Asplenium tricholepis, Asplenium trichomanes L., Asplenium trichomanessubsp. inexpectans, Asplenium trichomanes subsp. quadrivalens, Aspleniumtrichomanes subsp. trichomanes, Asplenium trichomanes var. harovii,Asplenium trichomanes var. herbaceum, Asplenium trichomanes var. repens,Asplenium trichomanes var. viridissimum, Asplenium trichomanes-dentatumL., Asplenium trigonopterum, Asplenium trilobatum, Asplenium trilobum,Asplenium triphyllum, Asplenium triphyllum var. compactum, Aspleniumtriphyllum var. gracillimum, Asplenium triphyllum var. herbaceum,Asplenium tripteropus, Asplenium triquetrum, Asplenium truncorum,Asplenium tsaratananense, Asplenium tucumanense, Asplenium tuerckheimii,Asplenium tunquiniense, Asplenium ulbrichtii, Asplenium ultimum,Asplenium unilaterale, Asplenium unilaterale var. decurrens, Aspleniumunilaterale var. udum, Asplenium unilaterale var. unilaterale, Aspleniumuniseriale, Asplenium uropteron, Asplenium vagans, Aspleniumvareschianum, Asplenium variabile var. paucijugum, Asplenium variabilevar. variabile, Asplenium varians subsp. fimbriatum, Asplenium varians,Asplenium vastum, Asplenium venturae, Asplenium venulosum, Aspleniumverapax, Asplenium vesiculosum, Asplenium vespertinum, Aspleniumvillosum, Asplenium virens, Asplenium viride, Asplenium viridifrons,Asplenium virillae, Asplenium viviparioides, Asplenium viviparum,Asplenium viviparum var viviparum, Asplenium viviparum var. lineatu,Asplenium volubile, Asplenium vulcanicum, Asplenium wacketii, Aspleniumwagneri, Asplenium wallichianum, Asplenium warneckei, Aspleniumwilfordii, Asplenium williamsii, Asplenium wrightii, Aspleniumwrightioides, Asplenium wuliangshanense, Asplenium xianqianense,Asplenium xinjiangense, Asplenium xinyiense, Asplenium yelagagense,Asplenium yoshinagae, Asplenium yunnanense, Asplenium zamiifolium,Asplenium zanzibaricum, Asplenium biscayneanum, Asplenium curtissii,Asplenium ebenoides, Asplenium herb-wagneri, Asplenium heteroresiliens,Asplenium kenzoi, Asplenium plenum, Asplenium wangii, andAsplenium×clermontiae, Asplenium×gravesii.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Blechnaceae, Genus Blechnum L.selected from but not limited to Blechnum amabile, Blechnumappendiculatum, Blechnum articulatum, Blechnum australe, Blechnumaustrobrasilianum, Blechnum binervatum, Blechnum blechnoides, Blechnumbrasiliense, Blechnum capense, Blechnum cartilagineum, Blechnumcastaneum, Blechnum chambersii, Blechnum chilense, Blechnum colensoi,Blechnum contiguum, Blechnum cordatum, Blechnum coriaceum, Blechnumdiscolor, Blechnum doodioides, Blechnum durum, Blechnum eburneum,Blechnum ensiforme, Blechnum filiforme, Blechnum fluviatile, Blechnumfragile, Blechnum fraseri, Blechnum fullagari, Blechnum gibbum, Blechnumglandulosum, Blechnum gracile, Blechnum hancockii, Blechnum hastatum,Blechnum howeanum, Blechnum indicum, Blechnum kunthianum, Blechnumlaevigatum, Blechnum loxense, Blechnum magellanicum, Blechnummembranaceum, Blechnum microbasis, Blechnum microphyllum, Blechnummilnei, Blechnum minus, Blechnum mochaenum, Blechnum montanum, Blechnummoorei, Blechnum moritzianum, Blechnum nigrum, Blechnum niponicum,Blechnum norfolkianum, Blechnum novae-zelandiae, Blechnum nudum,Blechnum obtusatum, Blechnum occidentale, Blechnum oceanicum, Blechnumorientale, Blechnum patersonii, Blechnum penna-marina, Blechnumpolypodioides, Blechnum procerum, Blechnum punctulatum, Blechnumsampaioanum, Blechnum schiedeanum, Blechnum schomburgkii, Blechnumserrulatum, Blechnum simillimum, Blechnum spicant, Blechnumstipitellatum, Blechnum tabulare, Blechnum triangularifolium, Blechnumvieillardii, Blechnum vulcanicum, Blechnum wattsii, Blechnum whelanii,and Blechnum wurunuran.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Dryopteridaceae GenusAcrophorus, Genus Acrorumohra, Genus Anapausia, Genus Arachniodes, GenusBolbitis, Genus Ctenitis, Genus Cyclodium, Genus Cyrtogonellum, GenusCyrtomidictyum, Genus Cyrtomium, Genus Diacalpe, Genus Didymochlaena,Genus Dryopsis, Genus Dryopteris, Genus Elaphoglossum, GenusHypodematium, Genus Lastreopsis, Genus Leptorumohra, Genus Leucostegia,Genus Lithostegia, Genus Lomagramma, Genus Maxonia, Genus Megalastrum,Genus Olfersia, Genus Peranema, Genus Phanerophlebia, GenusPhanerophlebiopsis, Genus Polybotrya, Genus Polystichopsis, GenusPolystichum, Genus Rumohra, Genus Sorolepidium, Genus Stigmatopteris orGenus Teratophyllum. In some embodiments the PtIP-65 polypeptide isderived from a fern species in the Order Polypodiales, FamilyDryopteridaceae, Genus Bolbitis, selected from but not limited toBolbitis acrostichoides, Bolbitis aliena, Bolbitis angustipinna,Bolbitis appendiculata, Bolbitis auriculata, Bolbitis bemoullii,Bolbitis bipinnatifida, Bolbitis cadieri, Bolbitis christensenfi,Bolbitis confertifolia, Bolbitis costata, Bolbitis crispatula, Bolbitisfluviatilis, Bolbitis gaboonensis, Bolbitis gemmifera, Bolbitishainanensis, Bolbitis hastata, Bolbitis hekouensis, Bolbitis hemiotis,Bolbitis heteroclita, Bolbitis heudelotii, Bolbitis humblotii, Bolbitisinterlineata, Bolbitis latipinna, Bolbitis laxireticulate, Bolbitislindigfi, Bolbitis lonchophora, Bolbitis longiflagellata, Bolbitismajor, Bolbitis media, Bolbitis nicotianifolia, Bolbitis nodiflora,Bolbitis novoguineensis, Bolbitis oligarchica, Bolbitis palustris,Bolbitis pandurifolia, Bolbitis pergamentacea, Bolbitis portoricensis,Bolbitis presliana, Bolbitis quoyana, Bolbitis rawsonfi, Bolbitisrepanda, Bolbitis rhizophylla, Bolbitis riparia, Bolbitis rivularis,Bolbitis sagenioides, Bolbitis salicina, Bolbitis scalpturata, Bolbitisscandens, Bolbitis semicordata, Bolbitis semipinnatifida, Bolbitisserrata, Bolbitis serratifolia, Bolbitis simplex, Bolbitis sinensis,Bolbitis singaporensis, Bolbitis sinuata, Bolbitis subcordata, Bolbitissubcrenata, Bolbitis taylorii, Bolbitis tibetica, Bolbitis tonkinensis,Bolbitis umbrosa, Bolbitis vanuaensis, and Bolbitis virens.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis. In some embodiments the PtIP-65 polypeptide is derived froma fern species in the Order Polypodiales, Family Lomariopsidaceae, GenusNephrolepis is selected from but not limited to Nephrolepis abrupta,Nephrolepis acuminata, Nephrolepis acutifolia, Nephrolepis arida,Nephrolepis arthropteroides, Nephrolepis biserrata var. auriculata,Nephrolepis brownii, Nephrolepis celebica, Nephrolepis clementis,Nephrolepis cordifolia, Nephrolepis davalliae, Nephrolepis davallioides,Nephrolepis dayakorum, Nephrolepis delicatula, Nephrolepisdicksonioides, Nephrolepis duffii, Nephrolepis exaltate ssp. exaltatessp. Hawaiiensis, Nephrolepis falcate, Nephrolepis falciformis,Nephrolepis glabra, Nephrolepis hirsutula, Nephrolepis humatoides,Nephrolepis iridescens, Nephrolepis kurotawae, Nephrolepis laurifolia,Nephrolepis lauterbachii, Nephrolepis lindsayae, Nephrolepis multifida,Nephrolepis multiflora, Nephrolepis niphoboloides, Nephrolepisobliterate, Nephrolepis paludosa, Nephrolepis pectinate, Nephrolepispendula, Nephrolepis persicifolia, Nephrolepis pickelii, Nephrolepispilosula, Nephrolepis pubescens, Nephrolepis pumicicola, Nephrolepisradicans, Nephrolepis rivularis, Nephrolepis rosenstockii, Nephrolepissaligna, Nephrolepis schlechteri, Nephrolepis serrate, Nephrolepisthomsoni, Nephrolepis undulate var. aureoglandulosa, Nephrolepis×averyi,Nephrolepis×copelandii, and Nephrolepis×medlerae.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusCampyloneurum, Genus Drynaria, Genus Lepisorus, Genus Microgramma, GenusMicrosorum, Genus Neurodium, Genus Niphidium, Genus Pecluma M.G., GenusPhlebodium, Genus Phymatosorus, Genus Platycerium, Genus Pleopeltis,Genus Polypodium L.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPolypodium L. In some embodiments the PtIP-65 polypeptide is derivedfrom a fern species in the Order Polypodiales, Family Polypodiaceae,Genus Polypodium L. selected from but not limited to Polypodiumabsidatum, Polypodium acutifolium, Polypodium adiantiforme, Polypodiumaequale, Polypodium affine, Polypodium albidopaleatum, Polypodiumalcicorne, Polypodium alfarii, Polypodium alfredii, Polypodium alfrediivar. curtii, Polypodium allosuroides, Polypodium alsophilicola,Polypodium amamianum, Polypodium amoenum, Polypodium amorphum,Polypodium anetioides, Polypodium anfractuosum, Polypodium anguinum,Polypodium angustifolium f. remotifolia, Polypodium angustifolium var.amphostenon, Polypodium angustifolium var. heterolepis, Polypodiumangustifolium var. monstrosa, Polypodium angustipaleatum, Polypodiumangustissimum, Polypodium anisomeron var. pectinatum, Polypodiumantioquianum, Polypodium aoristisorum, Polypodium apagolepis, Polypodiumapicidens, Polypodium apiculatum, Polypodium apoense, Polypodiumappalachianum, Polypodium appressum, Polypodium arenarium, Polypodiumargentinum, Polypodium argutum, Polypodium armatum, Polypodiumaromaticum, Polypodium aspersum, Polypodium assurgens, Polypodium atrum,Polypodium auriculatum, Polypodium balaonense, Polypodium balliviani,Polypodium bamleri, Polypodium bangii, Polypodium bartlettii, Polypodiumbasale, Polypodium bemoullii, Polypodium biauritum, Polypodium bifrons,Polypodium blepharodes, Polypodium bolivari, Polypodium bolivianum,Polypodium bolobense, Polypodium bombycinum, Polypodium bombycinum var.insularum, Polypodium bradeorum, Polypodium bryophilum, Polypodiumbryopodum, Polypodium buchtienii, Polypodium buesii, Polypodiumbulbotrichum, Polypodium caceresii, Polypodium californicum f.brauscombii, Polypodium californicum f. parsonsiae, Polypodiumcalifornicum, Polypodium calophlebium, Polypodium calvum, Polypodiumcamptophyllarium var. abbreviatum, Polypodium capitellatum, Polypodiumcarpinterae, Polypodium chachapoyense, Polypodium chartaceum, Polypodiumchimantense, Polypodium chiricanum, Polypodium choquetangense,Polypodium christensenii, Polypodium christii, Polypodium chrysotrichum,Polypodium ciliolepis, Polypodium cinerascens, Polypodium collinsii,Polypodium colysoides, Polypodium confluens, Polypodium conforme,Polypodium confusum, Polypodium congregatifolium, Polypodium connellii,Polypodium consimile var. bourgaeanum, Polypodium consimile var. minor,Polypodium conterminans, Polypodium contiguum, Polypodium cookii,Polypodium coriaceum, Polypodium coronans, Polypodium costaricense,Polypodium costatum, Polypodium crassifolium f. angustissimum,Polypodium crassifolium var. longipes, Polypodium crassulum, Polypodiumcraterisorum, Polypodium cryptum, Polypodium crystalloneuron, Polypodiumcucullatum var. planum, Polypodium cuencanum, Polypodium cumingianum,Polypodium cupreolepis, Polypodium curranii, Polypodium curvans,Polypodium cyathicola, Polypodium cyathisorum, Polypodium cyclocolpon,Polypodium daguense, Polypodium damunense, Polypodium dareiformioides,Polypodium dasypleura, Polypodium decipiens, Polypodium decorum,Polypodium delicatulum, Polypodium deltoideum, Polypodium demeraranum,Polypodium denticulatum, Polypodium diaphanum, Polypodium dilatatum,Polypodium dispersum, Polypodium dissectum, Polypodium dissimulans,Polypodium dolichosorum, Polypodium dolorense, Polypodiumdonnell-smithii, Polypodium drymoglossoides, Polypodium ebeninum,Polypodium eggersii, Polypodium elmeri, Polypodium elongatum, Polypodiumenterosoroides, Polypodium erubescens, Polypodium erythrolepis,Polypodium erythrotrichum, Polypodium eurybasis, Polypodium eurybasisvar. villosum, Polypodium exornans, Polypodium falcoideum, Polypodiumfallacissimum, Polypodium farinosum, Polypodium faucium, Polypodiumfeel, Polypodium ferrugineum, Polypodium feuillei, Polypodium firmulum,Polypodium firmum, Polypodium flaccidum, Polypodium flagellare,Polypodium flexuosum, Polypodium flexuosum var. ekmanii, Polypodiumforbesii, Polypodium formosanum, Polypodium fraxinifolium subsp.articulatum, Polypodium fraxinifolium subsp. luridum, Polypodiumfructuosum, Polypodium fucoides, Polypodium fulvescens, Polypodiumgaleottii, Polypodium glaucum, Polypodium glycyrrhiza, Polypodiumgracillimum, Polypodium gramineum, Polypodium grandifolium, Polypodiumgratum, Polypodium graveolens, Polypodium griseo-nigrum, Polypodiumgriseum, Polypodium guttatum, Polypodium haalilioanum, Polypodiumhammatisorum, Polypodium hancockii, Polypodium haplophlebicum,Polypodium harrisii, Polypodium hastatum var. simplex, Polypodiumhawaiiense, Polypodium heanophyllum, Polypodium helleri, Polypodiumhemionitidium, Polypodium henryi, Polypodium herzogii, Polypodiumhesperium, Polypodium hessii, Polypodium hombersleyi, Polypodiumhostmannii, Polypodium humile, Polypodium hyalinum, Polypodium iboense,Polypodium induens var. subdentatum, Polypodium insidiosum, Polypodiuminsigne, Polypodium intermedium subsp. masafueranum var. obtuseserratum,Polypodium intramarginale, Polypodium involutum, Polypodium itatiayense,Polypodium javanicum, Polypodium juglandifolium, Polypodium kaniense,Polypodium knowltoniorum, Polypodium kyimbilense, Polypodiuml'herminieri var. costaricense, Polypodium lachniferum f. incurvata,Polypodium lachniferum var. glabrescens, Polypodium lachnopus,Polypodium lanceolaturn var. cornplanatum, Polypodium lanceolaturn var.trichophorum, Polypodium latevagans, Polypodium laxifrons, Polypodiumlaxifrons var. lividum, Polypodium lehmannianum, Polypodium leiorhizum,Polypodium leptopodon, Polypodium leuconeuron var. angustifolia,Polypodium leuconeuron var. latifolium, Polypodium leucosticta,Polypodium limulum, Polypodium lindigii, Polypodium lineatum, Polypodiumlomarioides, Polypodium longifrons, Polypodium loretense, Polypodiumloriceum var. umbraticum, Polypodium loriforme, Polypodium loxogramme f.gigas, Polypodium ludens, Polypodium luzonicum, Polypodium lycopodioidesf. obtusum, Polypodium lycopodioides L., Polypodium macrolepis,Polypodium macrophyllum, Polypodium macrosorum, Polypodiummacrosphaerum, Polypodium maculosum, Polypodium madrense, Polypodiummanmeiense, Polypodium margaritiferum, Polypodium maritimum, Polypodiummartensii, Polypodium mayoris, Polypodium megalolepis, Polypodiummelanotrichum, Polypodium menisciifolium var. pubescens, Polypodiummeniscioides, Polypodium merrillii, Polypodium mettenii, Polypodiummexiae, Polypodium microsorum, Polypodium militare, Polypodium minimum,Polypodium minusculum, Polypodium mixtum, Polypodium mollendense,Polypodium mollissimum, Polypodium moniliforme var. minus, Polypodiummonoides, Polypodium monticola, Polypodium montigenum, Polypodiummoritzianum, Polypodium moultonii, Polypodium multicaudatum, Polypodiummultilineatum, Polypodium multisorum, Polypodium munchii, Polypodiummuscoides, Polypodium myriolepis, Polypodium myriophyllum, Polypodiummyriotrichum, Polypodium nematorhizon, Polypodium nemorale, Polypodiumnesioticum, Polypodium nigrescentium, Polypodium nigripes, Polypodiumnigrocinctum, Polypodium nimbatum, Polypodium nitidissimum, Polypodiumnitidissimum var. latior, Polypodium nubrigenum, Polypodium oligolepis,Polypodium oligosorum, Polypodium oligosorum, Polypodium olivaceum,Polypodium olivaceum var. elatum, Polypodium oodes, Polypodiumoosphaerum, Polypodium oreophilum, Polypodium omatissimum, Polypodiumornatum, Polypodium ovatum, Polypodium oxylobum, Polypodium oxypholis,Polypodium pakkaense, Polypodium pallidum, Polypodium palmatopedatum,Polypodium palmeri, Polypodium panamense, Polypodium parvum, Polypodiumpatagonicum, Polypodium paucisorum, Polypodium pavonianum, Polypodiumpectinatum var. caliense, Polypodium pectinatum var. hispidum,Polypodium pellucidum, Polypodium pendulum var. boliviense, Polypodiumpercrassum, Polypodium perpusillum, Polypodium peruvianum var.subgibbosum, Polypodium phyllitidis var. elongatum, Polypodiumpichinchense, Polypodium pilosissimum, Polypodium pilosissimum var.glabriusculum, Polypodium pilossimum var. tunguraquensis, Polypodiumpityrolepis, Polypodium platyphyllum, Polypodium playfairii, Polypodiumplebeium var. cooperi, Polypodium plectolepidioides, Polypodiumpleolepis, Polypodium plesiosorum var.i, Polypodium podobasis,Polypodium podocarpum, Polypodium poloense, Polypodium polydatylon,Polypodium polypodioides var. aciculare, Polypodium polypodioides var.michauxianum, Polypodium praetermissum, Polypodium preslianum var.immersum, Polypodium procerum, Polypodium procerum, Polypodiumproductum, Polypodium productum, Polypodium prolongilobum, Polypodiumpropinguum, Polypodium proteus, Polypodium pruinatum, Polypodiumpseudocapillare, Polypodium pseudofratemum, Polypodium pseudonutans,Polypodium pseudoserratum, Polypodium pulcherrimum, Polypodiumpulogense, Polypodium pungens, Polypodium purpusii, Polypodium radicale,Polypodium randallii, Polypodium ratiborii, Polypodium reclinaturn,Polypodium recreense, Polypodium repens var. abruptum, Polypodiumrevolvens, Polypodium rhachipterygium, Polypodium rhomboideum,Polypodium rigens, Polypodium robustum, Polypodium roraimense,Polypodium roraimense, Polypodium rosei, Polypodium rosenstockii,Polypodium rubidum, Polypodium rudimentum, Polypodium rusbyi, Polypodiumsablanianum, Polypodium sarmentosum, Polypodium saxicola, Polypodiumschenckii, Polypodium schlechteri, Polypodium scolopendria, Polypodiumscolopendria, Polypodium scolopendrium, Polypodium scouleri, Polypodiumscutulatum, Polypodium segregatum, Polypodium semihirsutum, Polypodiumsemihirsutum var. fuscosetosum, Polypodium senile var. minor, Polypodiumsericeolanatum, Polypodium serraeforme, Polypodium serricula, Polypodiumsesquipedala, Polypodium sessilifolium, Polypodium setosum var. calvum,Polypodium setulosum, Polypodium shaferi, Polypodium sibomense,Polypodium siccum, Polypodium simacense, Polypodium simulans, Polypodiumsingeri, Polypodium sinicum, Polypodium sintenisii, Polypodium skutchii,Polypodium sloanei, Polypodium sodiroi, Polypodium sordidulum,Polypodium sordidum, Polypodium sphaeropteroides, Polypodium sphenodes,Polypodium sprucei, Polypodium sprucei var. furcativenosa, Polypodiumsteirolepis, Polypodium stenobasis, Polypodium stenolepis, Polypodiumstenopterum, Polypodium subcapillare, Polypodium subflabelliforme,Polypodium subhemionitidium, Polypodium subinaequale, Polypodiumsubintegrum, Polypodium subspathulatum, Polypodium subtile, Polypodiumsubvestitum, Polypodium subviride, Polypodium superficiale var.attenuatum, Polypodium superficiale var. chinensis, Polypodiumsursumcurrens, Polypodium tablazianurn, Polypodium taenifolium,Polypodium tamandarei, Polypodium tatei, Polypodium tenuiculum var.acrosora, Polypodium tenuiculum var. brasiliense, Polypodium tenuilore,Polypodium tenuinerve, Polypodium tepuiense, Polypodium teresae,Polypodium tetragonum var. incompletum, Polypodium thysanolepis var.bipinnatifidum, Polypodium thyssanolepis, var. thyssanolepis, Polypodiumthyssanolepsi, Polypodium tobagense, Polypodium trichophyllum,Polypodium tridactylum, Polypodium tridentatum, Polypodium trifurcaturnvar. brevipes, Polypodium triglossum, Polypodium truncatulum, Polypodiumtruncicola var. major, Polypodium truncicola var. minor, Polypodiumtuberosum, Polypodium tunguraguae, Polypodium turquinurn, Polypodiumturrialbae, Polypodium ursipes, Polypodium vegans, Polypodiumvaldealaturn, Polypodium versteegii, Polypodium villagranii, Polypodiumvirginianum f. cambroideum, Polypodium virginianurn f. peraferens,Polypodium vittarioides, Polypodium vulgare, Polypodium vulgare L.,Polypodium vulgare subsp. oreophilum, Polypodium vulgare var.acuminatum, Polypodium vulpinum, Polypodium williamsii, Polypodiumwobbense, Polypodium×fallacissimum-guttatum, Polypodium xantholepis,Polypodium xiphopteris, Polypodium yarumalense, Polypodium yungense, andPolypodium zosteriforme.

In some embodiments the PtIP-65 polypeptide is derived from a fernspecies in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium. In some embodiments the PtIP-65 polypeptide is derived froma fern species in the Order Polypodiales, Family Polypodiaceae, GenusPlatycerium selected from but not limited to Platycerium alcicorne,Platycerium andinum, Platycerium angolense, Platycerium bifurcatum,Platycerium coronarium, Platycerium elephantotis, Platycerium ellisii,Platycerium grande, Platycerium hillii, Platycerium holttumfi,Platycerium madagascariense, Platycerium quadridichotomum, Platyceriumridleyi, Platycerium sp. ES-2011, Platycerium stemaria, Platyceriumsuperbum, Platycerium veitchii, Platycerium wallichii, Platyceriumwandae, Platycerium wilhelminae-reginae, and Platycerium willinckii.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Division Lycophyta.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Division Lycophyta, Class Isoetopsida or Class Lycopodiopsida.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Class Isoetopsida Order Selaginales. In some embodiments the PtIP-65polypeptide is derived from a fern species in the Class Isoetopsida,Order Selaginales, Family Selaginellaceae. In some embodiments thePtIP-65 polypeptide is derived from a species in the Genus Selaginella.In some embodiments the PtIP-65 polypeptide is derived from aSelaginella species selected from but not limited to Selaginellaacanthonota, Selaginella apoda, Selaginella arbuscula, Selaginellaarenicola, Selaginella arizonica, Selaginella armata, Selaginellaasprella, Selaginella biformis, Selaginella bigelovii, Selaginellabraunii, Selaginella cinerascens, Selaginella cordifolia, Selaginelladeflexa, Selaginella delicatula, Selaginella densa, Selaginelladouglasii, Selaginella eatonii, Selaginella eclipes, Selaginellaeremophila, Selaginella erythropus, Selaginella flabellata, Selaginellahansenii, Selaginella heterodonta, Selaginella kraussiana, Selaginellakrugii, Selaginella laxifolia, Selaginella lepidophylla, Selaginellaleucobryoides, Selaginella ludoviciana, Selaginella mutica, Selaginellaoregana, Selaginella ovifolia, Selaginella pallescens, Selaginellaperuviana, Selaginella pilifera, Selaginella plana, Selaginella plumosa,Selaginella pulcherrima, Selaginella rupestris, Selaginella rupincola,Selaginella scopulorum, Selaginella selaginoides, Selaginella sibirica,Selaginella standleyi, Selaginella stellata, Selaginella subcaulescens,Selaginella substipitata, Selaginella tenella, Selaginella tortipila,Selaginella uliginosa, Selaginella umbrosa, Selaginella uncinata,Selaginella underwoodii, Selaginella utahensis, Selaginella victoriae,Selaginella viridissima, Selaginella wallacei, Selaginella watsonii,Selaginella weatherbiana, Selaginella willdenowii, Selaginella wrightiiand Selaginella×neomexicana.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Class Lycopodiopsida, Order Lycopodiales. In some embodiments thePtIP-65 polypeptide is derived from a fern species in the ClassLycopodiopsida, Order Lycopodiales Family Lycopodiaceae or FamilyHuperziaceae. In some embodiments the PtIP-65 polypeptide is derivedfrom a species in the Genus Austrolycopodium, Dendrolycopodium,Diphasiastrum, Diphasium, Huperzia, Lateristachys, Lycopodiastrum,Lycopodiella, Lycopodium, Palhinhaea, Pseudodiphasium,Pseudolycopodiella, Pseudolycopodium or Spinulum.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Genus Lycopodium. In some embodiments the PtIP-65 polypeptide isderived from a Lycopodium species selected from but not limited toLycopodium alpinum L., Lycopodium annotinum L., Lycopodium clavatum L.,Lycopodium complanatum L., Lycopodium dendroideum Michx., Lycopodiumdigitatum, Lycopodium xhabereri, Lycopodium hickeyi, Lycopodium×issleri,Lycopodium lagopus, Lycopodium obscurum L., Lycopodium phlegmaria L.,Lycopodium sabinifolium, Lycopodium sitchense, Lycopodium tristachyum,Lycopodium venustulum, Lycopodium venustulum var. venustulum, Lycopodiumvenustulum var. verticale, Lycopodium volubile and Lycopodium×zeilleri.

In some embodiments the PtIP-65 polypeptide is derived from a species inthe Genus Huperzia. In some embodiments the PtIP-65 polypeptide isderived from a species selected from but not limited to Huperziaappressa, Huperzia arctica, Huperzia attenuata, Huperzia australiana,Huperzia balansae, Huperzia billardierei, Huperzia brassii, Huperziacampiana, Huperzia capellae, Huperzia carinata, Huperzia cf. carinataARF000603, Huperzia cf. nummulariifolia ARF001140, Huperzia cf.phlegmaria ARF000717, Huperzia cf. phlegmaria ARF000771, Huperzia cf.phlegmaria ARF000785, Huperzia cf. phlegmaria ARF001007, Huperzia cf.phlegmaria ARF002568, Huperzia cf. phlegmaria ARF002703, Huperzia cf.phlegmaria Wikstrom 1998, Huperzia chinensis, Huperzia compacta,Huperzia crassa, Huperzia crispata, Huperzia cryptomeriana, Huperziacumingii, Huperzia dacrydioides, Huperzia dalhousieana, Huperziadichotoma, Huperzia emeiensis, Huperzia ericifolia, Huperzia eversa,Huperzia fargesii, Huperzia fordii, Huperzia funiformis, Huperziagoebellii, Huperzia haleakalae, Huperzia hamiltonii, Huperziaheteroclita, Huperzia hippuridea, Huperzia hippuris, Huperzia holstii,Huperzia horizontalis, Huperzia hunanensis, Huperzia hystrix, Huperzialindenii, Huperzia linifolia, Huperzia lockyeri, Huperzia lucidula,Huperzia mingcheensis, Huperzia miyoshiana, Huperzia nanchuanensis,Huperzia nummulariifolia, Huperzia obtusifolia, Huperziaophioglossoides, Huperzia petiolata, Huperzia phlegmaria, Huperziaphlegmarioides, Huperzia phyllantha, Huperzia pinifolia, Huperziapolydactyla, Huperzia prolifera, Huperzia reflexa, Huperziarosenstockiana, Huperzia rufescens, Huperzia salvinoides, Huperziasarmentosa, Huperzia selago, Huperzia serrata, Huperzia sieboldii,Huperzia somae, Huperzia squarrosa, Huperzia subulata, Huperziasutchueniana, Huperzia tauri, Huperzia taxifolia, Huperzia tenuis,Huperzia tetragona, Huperzia tetrasticha, Huperzia unguiculata, Huperziavaria, Huperzia verticillata and Huperzia wilsonii.

In some embodiments the PtIP-65 polypeptide comprises at least one ofthe amino acid sequence motifs as shown in Table 3.

TABLE 3 Motif 12 Motif 2 Motif 19 Motif 6 Motif 17 Motif 1 Motif 10PtIP-65Gc SEQ ID NO: 27 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Ha SEQ ID NO: 25 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Gd SEQ ID NO: 35 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Ge SEQ ID NO: 36 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Ga SEQ ID NO: 30 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Gb SEQ ID NO: 34 2-17 18-48 49-60 61-76 n.i. 77-119 n.i.PtIP-65Hj SEQ ID NO: 39 2-17 18-48 49-60 62-77 n.i. 78-120 n.i.PtIP-65Hk SEQ ID NO: 37 n.i. n.i. n.i. 63-77 n.i. n.i. n.i. PtIP-65HgSEQ ID NO: 40 82-97   98-128 n.i. 141-156 n.i. 157-199  n.i. PtIP-65HhSEQ ID NO: 38 41-56  57-87 n.i. 103-118 n.i. 119-161  n.i. PtIP-65Fa SEQID NO: 41 1-16 18-48 n.i. 67-82 50-65 83-125 n.i. PtIP-65Fb SEQ ID NO:42 1-16 18-48 n.i. 67-82 50-65 83-125 n.i. PtIP-65Ca SEQ ID NO: 29 0-1516-46 n.i. 65-80 48-63 81-123 n.i. PtIP-65Aa SEQ ID NO: 22 0-15 16-46n.i. 65-80 48-63 81-123 n.i. PtIP-65Ba SEQ ID NO: 24 0-15 16-46 n.i.65-80 48-63 81-123 n.i. PtIP-65Bb SEQ ID NO: 26 0-15 16-46 n.i. 65-8048-63 81-123 n.i. PtIP-65He SEQ ID NO: 32 n.i. 16-46 47-58 60-75 n.i.n.i. 86-127 PtIP-65Hf SEQ ID NO: 33 n.i. 22-52 53-64 66-81 n.i. n.i.93-134 PtIP-65Hb SEQ ID NO: 31 n.i. 21-51 52-63 64-79 n.i. n.i. 95-136PtIP-65Hc SEQ ID NO: 23 n.i. 23-53 54-65 66-81 n.i. n.i. 97-138PtIP-65Hd SEQ ID NO: 28 n.i. 23-53 54-65 66-81 n.i. n.i. 98-139 Motif 16Motif 3 Motif 15 Motif 27 Motif 7 Motif 11 Motif 21 PtIP-65Gc SEQ ID NO:27 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Ha SEQ ID NO:25 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Gd SEQ ID NO:35 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Ge SEQ ID NO:36 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Ga SEQ ID NO:30 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Gb SEQ ID NO:34 120-128 129-159 161-172 n.i. 175-204 n.i. n.i. PtIP-65Hj SEQ ID NO:39 121-129 130-160 162-173 n.i. 176-207 n.i. n.i. PtIP-65Hk SEQ ID NO:37 n.i. 133-163 164-175 n.i. 179-208 n.i. n.i. PtIP-65Hg SEQ ID NO: 40200-208 209-239 241-252 n.i. n.i. n.i. n.i. PtIP-65Hh SEQ ID NO: 38162-170 n.i. 204-215 n.i. 217-246 n.i. n.i. PtIP-65Fa SEQ ID NO: 41126-134 135-165 167-178 n.i. 180-209 n.i. n.i. PtIP-65Fb SEQ ID NO: 42126-134 135-165 167-178 n.i. 180-209 n.i. n.i. PtIP-65Ca SEQ ID NO: 29124-132 133-163 n.i. n.i. n.i. 182-222 n.i. PtIP-65Aa SEQ ID NO: 22124-132 133-163 n.i. n.i. n.i. 182-222 n.i. PtIP-65Ba SEQ ID NO: 24124-132 133-163 n.i. n.i. n.i. 182-222 n.i. PtIP-65Bb SEQ ID NO: 26124-132 133-163 n.i. n.i. n.i. 182-222 n.i. PtIP-65He SEQ ID NO: 32 n.i.131-161 n.i. 169-177 178-207 n.i. 217-238 PtIP-65Hf SEQ ID NO: 33 n.i.137-167 n.i. 175-183 184-213 n.i. 223-244 PtIP-65Hb SEQ ID NO: 31 n.i.139-169 n.i. 177-185 186-215 n.i. 222-243 PtIP-65Hc SEQ ID NO: 23 n.i.141-171 n.i. 178-186 187-216 n.i. 225-246 PtIP-65Hd SEQ ID NO: 28 n.i.142-172 n.i. 179-187 188-217 n.i. 226-247 Motif 5 Motif 20 Motif 26Motif 23 Motif 18 Motif 4 PtIP-65Gc SEQ ID NO: 27 206-236 n.i. n.i.239-250 n.i. 251-300 PtIP-65Ha SEQ ID NO: 25 206-236 n.i. n.i. 239-250n.i. 251-300 PtIP-65Gd SEQ ID NO: 35 206-236 n.i. n.i. 239-250 n.i.251-300 PtIP-65Ge SEQ ID NO: 36 206-236 n.i. n.i. 239-250 n.i. 251-300PtIP-65Ga SEQ ID NO: 30 206-236 n.i. n.i. 239-250 n.i. 251-300 PtIP-65GbSEQ ID NO: 34 206-236 n.i. n.i. 239-250 n.i. 251-300 PtIP-65Hj SEQ IDNO: 39 207-237 n.i. n.i. 240-251 n.i. 252-301 PtIP-65Hk SEQ ID NO: 37210-240 n.i. n.i. n.i. n.i. n.i. PtIP-65Hg SEQ ID NO: 40 285-315 n.i.n.i. n.i. n.i. 330-379 PtIP-65Hh SEQ ID NO: 38 248-278 n.i. 307-322 n.i.n.i. n.i. PtIP-65Fa SEQ ID NO: 41 211-241 n.i. n.i. n.i. n.i. 256-305PtIP-65Fb SEQ ID NO: 42 211-241 n.i. n.i. n.i. n.i. 256-305 PtIP-65CaSEQ ID NO: 29 229-259 260-288 291-306 n.i. n.i. n.i. PtIP-65Aa SEQ IDNO: 22 229-259 260-288 290-305 n.i. n.i. n.i. PtIP-65Ba SEQ ID NO: 24n.i. 261-289 291-306 n.i. n.i. n.i. PtIP-65Bb SEQ ID NO: 26 229-259260-288 290-305 n.i. n.i. n.i. PtIP-65He SEQ ID NO: 32 n.i. n.i. n.i.n.i. 241-262 n.i. PtIP-65Hf SEQ ID NO: 33 n.i. n.i. n.i. n.i. 247-268n.i. PtIP-65Hb SEQ ID NO: 31 n.i. n.i. n.i. n.i. 245-266 n.i. PtIP-65HcSEQ ID NO: 23 n.i. n.i. V n.i. 248-269 n.i. PtIP-65Hd SEQ ID NO: 28 n.i.n.i. n.i. n.i. 249-270 n.i. Motif 24 Motif 13 Motif 22 Motif 9 Motif 14Motif 8 PtIP-65Gc SEQ ID NO: 27 n.i. 302-317 319-330 n.i. n.i. 332-353PtIP-65Ha SEQ ID NO: 25 n.i. 302-317 319-330 n.i. n.i. 332-353 PtIP-65GdSEQ ID NO: 35 n.i. 302-317 318-329 n.i. n.i. 331-352 PtIP-65Ge SEQ IDNO: 36 n.i. 302-317 318-329 n.i. n.i. 331-352 PtIP-65Ga SEQ ID NO: 30n.i. 302-317 318-329 n.i. n.i. 332-353 PtIP-65Gb SEQ ID NO: 34 n.i.302-317 318-329 n.i. n.i. 332-353 PtIP-65Hj SEQ ID NO: 39 n.i. 303-318n.i. n.i. n.i. n.i. PtIP-65Hk SEQ ID NO: 37 n.i. n.i. n.i. n.i. n.i.n.i. PtIP-65Hg SEQ ID NO: 40 n.i. n.i. n.i. n.i. n.i. n.i. PtIP-65Hh SEQID NO: 38 n.i. n.i. n.i. n.i. n.i. n.i. PtIP-65Fa SEQ ID NO: 41 n.i.n.i. n.i. n.i. n.i. n.i. PtIP-65Fb SEQ ID NO: 42 n.i. n.i. n.i. n.i.n.i. n.i. PtIP-65Ca SEQ ID NO: 29 310-340 n.i. n.i. n.i. n.i. n.i.PtIP-65Aa SEQ ID NO: 22 308-338 n.i. n.i. n.i. n.i. n.i. PtIP-65Ba SEQID NO: 24 309-339 n.i. n.i. n.i. n.i. n.i. PtIP-65Bb SEQ ID NO: 26308-338 n.i. n.i. n.i. n.i. n.i. PtIP-65He SEQ ID NO: 32 n.i. n.i. n.i.263-298 300-349 n.i. PtIP-65Hf SEQ ID NO: 33 n.i. n.i. n.i. 269-304 n.i.PtIP-65Hb SEQ ID NO: 31 n.i. n.i. n.i. 267-302 303-352 n.i. PtIP-65HcSEQ ID NO: 23 n.i. n.i. n.i. 270-305 306-355 n.i. PtIP-65Hd SEQ ID NO:28 n.i. n.i. n.i. 272-307 308-357 n.i.

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); and at least one aminoacid sequence motif selected from: i) an amino acid sequence motif 2 asrepresented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); and ii) an amino acid sequence motif 3 as representedby an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); and at least one amino acid sequence motif selectedfrom: i) an amino acid sequence motif 1 as represented by an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); ii) an amino acid sequence motif 5 as represented byan amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); iii) an amino acid sequence motif 12 as represented byan amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS](SEQ ID NO: 158); andiv) an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO:162).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); ii) an amino acid sequence motif 17 as represented byan amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); iii) anamino acid sequence motif 20 as represented by an amino acid sequence ofthe formula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO:166); iv) an amino acid sequence motif 24 as represented by an aminoacid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and v) an amino acid sequence motif 26 as representedby an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); anamino acid sequence motif 11 as represented by an amino acid sequence ofthe formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); an aminoacid sequence motif 20 as represented by an amino acid sequence of theformula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166);an amino acid sequence motif 24 as represented by an amino acid sequenceof the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and an amino acid sequence motif 26 as represented byan amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 as represented by an amino acid sequence ofthe formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ IDNO: 152); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 11 as represented by anamino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 20 as represented by anamino acid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF](SEQ ID NO: 166); an aminoacid sequence motif 26 as represented by an amino acid sequence of theformula L[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQID NO: 171) and an amino acid sequence motif 24 as represented by anamino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 as represented by an amino acid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); ii) an amino acid sequence motif 13 as represented by an aminoacid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO:159); iii) an amino acid sequence motif 19 as represented by an aminoacid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); iv)an amino acid sequence motif 22 as represented by an amino acid sequenceof the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); andv) an amino acid sequence motif 23 as represented by an amino acidsequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 as represented by anamino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); anamino acid sequence motif 8 as represented by an amino acid sequence ofthe formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); an amino acid sequence motif 13 as represented by an aminoacid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO:159); an amino acid sequence motif 19 as represented by an amino acidsequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 22 as represented by an amino acid sequence ofthe formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); an aminoacid sequence motif 23 as represented by an amino acid sequence of theformula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169); and at least one amino acidsequence motif selected from: i) an amino acid sequence motif 4 asrepresented by an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); ii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iii) an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 as represented by an amino acid sequence ofthe formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 as represented by an amino acid sequence ofthe formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif selected from: an amino acid sequence motif 1 asrepresented by an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 as represented by an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 23 as represented by anamino acid sequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169);an amino acid sequence motif 4 as represented by an amino acid sequenceof the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ IDNO: 159); an amino acid sequence motif 22 as represented by an aminoacid sequence of the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ IDNO: 168); and an amino acid sequence motif 8 as represented by an aminoacid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 as represented by an amino acid sequence ofthe formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 10 as represented by an amino acid sequence of theformula[WPY][DTN][HNV][VI][AG]R[VI][RW]F[SQ]TS[RL][GL]R[TI]FE[YF]G[SM][ST][RV][EQ]P[ST]G[QKR][WV][FIT][TE][VA]NVGSGVCVG[VM](SEQ ID NO: 156); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 27 as represented by anamino acid sequence of the formula T[TFPS]IL[EQS][EQH]LP[DA] (SEQ ID NO:172); an amino acid sequence motif 7 as represented by an amino acidsequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 21 as represented by anamino acid sequence of the formulaHL[TV][GA][NRS][AND]I[STL][IVL][ND][VILM][DTS]LGI[ND][TA]P[TK][VI]V[PA](SEQ ID NO: 167); an amino acid sequence motif 18 as represented by anamino acid sequence of the formulaG[TLP][AE]GAST[NTA]F[GQH]W[EQ]T[VA]R[TA]FPS[TIS]NA (SEQ ID NO: 164); anamino acid sequence motif 9 as represented by an amino acid sequence ofthe formula[IMV][QE]G[ST]I[AS][ND]L[TI]V[SRT]T[NAD][EA]YSVW[CG]H[IV]SDT[LVI]APAQ[SL][LI][PI][KS][HR][RASV]A (SEQ ID NO: 155); an amino acid sequence motif 14 asrepresented by an amino acid sequence of the formula[LW][VI]GEG[RS]I[TD][AN]L[PQ]CSA[NR]IQV[FIL]T[SD][GAS][GAD][NGY][NDG][LF]PF[AG][TS]F[ST]FPV[RGQ][LSV][LF]Y[DNS][GA][GR]AHS[DNQT]VQ[VI][LI](SEQ ID NO: 160).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST] (SEQ ID NO: 151); an amino acid sequence motif 12 asrepresented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 as represented by an amino acid sequence ofthe formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 as represented by an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163);ii) an amino acid sequence motif 4 as represented by an amino acidsequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); iii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iv) an amino acid sequence motif 15 as representedby an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 2 as represented by an amino acid sequence ofthe formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ IDNO: 152); an amino acid sequence motif 17 as represented by an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); an amino acid sequence motif selected from: an aminoacid sequence motif 1 as represented by an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 as represented by anamino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 as represented by an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); and an amino acid sequence motif 4 as represented byan amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); and ii) an amino acid sequence motif 3 having at least90% identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); and at least one amino acid sequence motif selectedfrom: i) an amino acid sequence motif 1 having at least 90% identity toan amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); ii) an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); iii) an amino acid sequence motif 12 having at least90% identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);and iv) an amino acid sequence motif 16 having at least 90% identity toan amino acid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR](SEQ ID NO: 162).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 11 having at least 90% identity to an aminoacid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); ii) an amino acid sequence motif 17 having at least90% identity to an amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); iii) anamino acid sequence motif 20 having at least 90% identity to an aminoacid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166); iv) anamino acid sequence motif 24 having at least 90% identity to an aminoacid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and v) an amino acid sequence motif 26 having at least90% identity to an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); an amino acid sequence motif 11 having at least 90% identityto an amino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 17 having at least 90%identity to an amino acid sequence of the formulaD[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); an aminoacid sequence motif 20 having at least 90% identity to an amino acidsequence of the formula DT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF](SEQ ID NO: 166); an amino acid sequence motif 24 having at least 90%identity to an amino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170); and an amino acid sequence motif 26 having at least90% identity to an amino acid sequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula [LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS](SEQ ID NO: 158); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 17 having at least 90% identity to an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); anamino acid sequence motif selected from: an amino acid sequence motif 1having at least 90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 11 having at least 90%identity to an amino acid sequence of the formulaK[ST]F[NT]L[QGL]N[TA]A[DHN][QH][DE]D[PQ]SST[EQ][QK]LAW[EQ]LADEP[SR]FDNV[SF][HL][ST]W[SI]LSN(SEQ ID NO: 157); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 20 having at least 90%identity to an amino acid sequence of the formulaDT[QE]SSSLSSSSSL[LS]LPWSR[NS]GSLLPS[EKQ]S[LF] (SEQ ID NO: 166); an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SV]AL[PT][YFR][RDGQ]G[NDIT][VAL][NSR][GDN]L[PFRS]F[NDET] (SEQ ID NO:171) and an amino acid sequence motif 24 having at least 90% identity toan amino acid sequence of the formula[AQ][RV][VT]TT[ST][GH][GS][TR]FSF[SP]GL[QHK]G[AFLR][FY]TG[QE]S[HY][VA][IP][AD][IV]T[TAS]Q(SEQ ID NO: 170).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158); anamino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 8 having at least 90% identity to an aminoacid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); ii) an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO: 159); iii) an amino acidsequence motif 19 having at least 90% identity to an amino acid sequenceof the formula [TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQID NO: 165); iv) an amino acid sequence motif 22 having at least 90%identity to an amino acid sequence of the formula[KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); and v) an amino acidsequence motif 23 having at least 90% identity to an amino acid sequenceof the formula Y[TA][ST]TQEESRTLT (SEQ ID NO: 169).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); an amino acid sequence motif 8 having at least 90% identity toan amino acid sequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154); an amino acid sequence motif 13 having at least 90% identityto an amino acid sequence of the formula GVDYTSVDI[TV][NS][TQ]GTR[AD](SEQ ID NO: 159); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 22 having at least 90% identity to an aminoacid sequence of the formula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ IDNO: 168); an amino acid sequence motif 23 having at least 90% identityto an amino acid sequence of the formula Y[TA][ST]TQEESRTLT (SEQ ID NO:169); and at least one amino acid sequence motif selected from: i) anamino acid sequence motif 4 having at least 90% identity to an aminoacid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); ii) an amino acid sequence motif 7 having at least 90%identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iii) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula [LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS](SEQ ID NO: 158); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO:152); an amino acid sequence motif selected from: an amino acid sequencemotif 1 having at least 90% identity to an amino acid sequence of theformulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 having at least 90% identity to an amino acid sequenceof the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 23 having at least 90%identity to an amino acid sequence of the formula Y[TA][ST]TQEESRTLT(SEQ ID NO: 169); an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TV][NS][TQ]GTR[AD] (SEQ ID NO: 159); an amino acid sequencemotif 22 having at least 90% identity to an amino acid sequence of theformula [KLS][DKN][EHQ]VEV[EQ]A[TV][DE]QQ (SEQ ID NO: 168); and an aminoacid sequence motif 8 having at least 90% identity to an amino acidsequence of the formula[QG][EV][GK]D[HQ][NS]VQP[ND]KE[VA][EKQ]E[CRS][KT][LV]LF[AIT]E (SEQ IDNO: 154).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTN][DS][TSN]P[AHYRIAT][TVL][FYC]G[TS][APV] (SEQ ID NO: 165); anamino acid sequence motif 6 having at least 90% identity to an aminoacid sequence of the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO:152); an amino acid sequence motif 10 having at least 90% identity to anamino acid sequence of the formula[WPY][DTN][HNV][VI][AG]R[VI][RW]F[SQ]TS[RL][GL]R[TI]FE[YF]G[SM][ST][RV][EQ]P[ST]G[QKR][WV][FIT][TE][VA]NVGSGVCVG[VM](SEQ ID NO: 156); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 27 having at least 90%identity to an amino acid sequence of the formulaT[TFPS]IL[EQS][EQH]LP[DA] (SEQ ID NO: 172); an amino acid sequence motif7 having at least 90% identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 21 having at least 90%identity to an amino acid sequence of the formulaHL[TV][GA][NRS][AND]I[STL][IVL][ND][VILM][DTS]LGI[ND][TA]P[TK][VI]V[PA](SEQ ID NO: 167); an amino acid sequence motif 18 having at least 90%identity to an amino acid sequence of the formulaG[TLP][AE]GAST[NTA]F[GQH]W[EQ]T[VA]R[TA]FPS[TIS]NA (SEQ ID NO: 164); anamino acid sequence motif 9 having at least 90% identity to an aminoacid sequence of the formula[IMV][QE]G[ST]I[AS][ND]L[TI]V[SRT]T[NAD][EA]YSVW[CG]H[IV]SDT[LVI]APAQ[SL][LI][PI][KS][HR][RASV]A (SEQ ID NO: 155); an amino acid sequence motif 14 havingat least 90% identity to an amino acid sequence of the formula[LW][VI]GEG[RS]I[TD][AN]L[PQ]CSA[NR]IQV[FIL]T[SD][GAS][GAD][NGY][NDG][LF]PF[AG][TS]F[ST]FPV[RGQ][LSV][LF]Y[DNS][GA][GR]AHS[DNQT]VQ[VI][LI](SEQ ID NO: 160).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence ofGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); an amino acid sequence motif 12 having at least 90%identity to an amino acid sequence of the formula[LM][AVY][QL][TV][P][TV][P][VG][YT][VL]IGGQGG[SN][AQS] (SEQ ID NO: 158);an amino acid sequence motif 16 having at least 90% identity to an aminoacid sequence of the formula [AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ IDNO: 162); and at least one amino acid sequence motif selected from: i)an amino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of the formula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE(SEQ ID NO: 163); ii) an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150); iii) an amino acid sequence motif 7 having at least90% identity to an amino acid sequence of the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); and iv) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula [LM][AVY][QL][TV][PI][VG][YT][VL]IGGQGG[SN][AQS](SEQ ID NO: 158); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STR]Y[DY][GQ][GSE][TR][DN]G[RK]V[LM][QTR][RK]I[GR]V[WY][AR][GA][ET][SW][RQ][IL][KR][AG]I[RES]VW[LM](SEQ ID NO: 148); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GL][SE]YSE[FY]TFA[DET]GERIT (SEQ ID NO: 152); an amino acid sequencemotif 17 having at least 90% identity to an amino acid sequence of theformula D[GE][VA][QEA][KQR][TL]FGDP[SH][RD]P[APS]GE (SEQ ID NO: 163); anamino acid sequence motif selected from: an amino acid sequence motif 1having at least 90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SA]G[WAG]I[RY]F[YER]T[TNS][TQ][GN][GR][SRT]F[FD][FPH][GK]M[TY]SWG[LK][KQ]TEYP[IVQ]DV(SEQ ID NO: 147); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVS]SG[ILV][CW][VA]G[IVLF][TILMR] (SEQ ID NO: 162); an amino acidsequence motif 3 having at least 90% identity to an amino acid sequenceof the formulaGRA[NS][SV]D[IVL][DN][SA]LG[FV][VML]FLR[PTS]I[QA]S[AVCS]R[LM]I[NS]V[QS]YPTL(SEQ ID NO: 149); an amino acid sequence motif 15 having at least 90%identity to an amino acid sequence of the formula[LF][ED][QT][AQ]GI[IVST]P[VITQ][TA]L[DK] (SEQ ID NO: 161); an amino acidsequence motif 7 having at least 90% identity to an amino acid sequenceof the formula[NT]D[SK]NN[AD][GDS]S[IE][PS][KL]NW[TV][FL][SA]GSR[TQ]VT[ITV]SS[ST]W[STR][LT]T(SEQ ID NO: 153); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQ][ILF][EIT][ATV][HT][AS]S[VI][TS]V[QES]A[GR]IP[AS][VL]AEV[SN]G[EV][FV]GW[SEQ][VI]S[VA][ST](SEQ ID NO: 151); and an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NG][QV]SGTL[ES][PA]G[QE][WS]I[SH][LI][QK]A[TL]TR[RK]G[TL]I[TS][LV]P[YF]Q[GA][TS][MI][EQ][IV]TL[QKL][SN]G[TD][VI]F[QR]Y[PA][IL][SK][SG][MQ]Y(SEQ ID NO: 150).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); and at least oneamino acid sequence motif selected from: i) an amino acid sequence motif2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); and ii) an amino acid sequence motif 3 as representedby an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); and at least one amino acid sequencemotif selected from: i) an amino acid sequence motif 1 as represented byan amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); ii) an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); iii) anamino acid sequence motif 12 as represented by an amino acid sequence ofthe formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); and iv) an amino acid sequence motif 16 as representedby an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); ii) anamino acid sequence motif 17 as represented by an amino acid sequence ofthe formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); iii) an amino acid sequence motif 20 as represented byan amino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); iv) an amino acid sequence motif 24 as represented by an aminoacid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and v) an aminoacid sequence motif 26 as represented by an amino acid sequence of theformulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 11 as represented by an amino acid sequence of theformulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 17 as represented by an amino acid sequence of theformula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ IDNO: 189); an amino acid sequence motif 20 as represented by an aminoacid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 24 as represented by an amino acidsequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and an aminoacid sequence motif 26 as represented by an amino acid sequence of theformulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 11 asrepresented by an amino acid sequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 5 as represented by an amino acid sequence of theformula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 20 as represented by an amino acid sequence of theformula DT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQID NO: 192); an amino acid sequence motif 26 as represented by an aminoacid sequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197) and an amino acid sequence motif 24 as represented byan amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 as represented by an amino acid sequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); ii) an amino acid sequence motif 13 as represented byan amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); iii) an aminoacid sequence motif 19 as represented by an amino acid sequence of theformula [TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL](SEQ ID NO: 191); iv) an amino acid sequence motif 22 as represented byan amino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and v)an amino acid sequence motif 23 as represented by an amino acid sequenceof the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 8 as represented by an amino acid sequence of theformula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 22 as represented by an amino acidsequence of the formula [KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ(SEQ ID NO: 194); an amino acid sequence motif 23 as represented by anamino acid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195);and at least one amino acid sequence motif selected from: i) an aminoacid sequence motif 4 as represented by an amino acid sequence of theformulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); ii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iii) an amino acid sequence motif as representedby an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 as represented by an amino acidsequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO:178); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 as represented by an amino acid sequence ofthe formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177) an aminoacid sequence motif 23 as represented by an amino acid sequence of theformula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); an amino acid sequencemotif 4 as represented by an amino acid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); an amino acid sequence motif 13 as represented by anamino acid sequence of the formula GVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acid sequence motif 22 as represented by anamino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and anamino acid sequence motif 8 as represented by an amino acid sequence ofthe formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 as represented by anamino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 as represented by an amino acidsequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO:178); an amino acid sequence motif 10 as represented by an amino acidsequence of the formula[WPYF][DTNQES][HNVQIL][VIL][AG]R[VIL][RWKYH]F[SQTN]TS[RLKIV][GLIV]R[TISLV]FE[YFW]G[SMT][ST][RV][EQND]P[ST]G[QKRN][WVYIL][FITLVS][TESD][VAIL]NVGSGVCVG[VMIL] (SEQ ID NO: 182); an amino acid sequence motif 3 as represented byan amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 27 asrepresented by an amino acid sequence of the formulaT[TFPS]IL[EQSDNT][EQHDN]LP[DAE] (SEQ ID NO: 198); an amino acid sequencemotif 7 as represented by an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 21 as represented by anamino acid sequence of the formulaHL[TVSIL][GA][NRSQKT][ANDQE]I[STLIV][IVL][NDQE][VILM][DTSE]LGI[ND][TAS]P[TKSR][VIL]V[PA] (SEQ ID NO: 193); an amino acid sequencemotif 18 as represented by an amino acid sequence of the formulaG[TLPSIV][AED]GAST[NTAQS]F[GQHN]W[EQND]T[VAIL]R[TAS]FPS[TISLV]NA (SEQ IDNO: 190); an amino acid sequence motif 9 as represented by an amino acidsequence of the formula[IMVL][QEND]G[ST]I[AST][NDQE]L[TISLV]V[SRTK]T[NADQE][EAD]YSVW[CG]H[IVL]SDT[LVI]APAQ[SLTIV][LIV][PIVL][KSRT][HRK][RASVKTIL]A (SEQ ID NO: 181); anamino acid sequence motif 14 as represented by an amino acid sequence ofthe formula[LWIVF][VIL]GEG[RSKT]I[TDSE][ANQ]L[PQN]CSA[NRQK]IQV[FILV]T[SDTE][GAST][GADE][NGYQ][NDGQE][LFIV]PF[AG][TS]F[ST]FPV[RGQKN][LSVIT][LFIV]Y[DNSQET][GA][GRK]AHS[DNQTES]VQ[VIL][LIV] (SEQ ID NO: 186).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 as represented by an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 2 as represented by an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 as represented by anamino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 as represented by an aminoacid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 as represented by an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 as represented by an amino acid sequence of theformula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 as represented by anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 as represented by an amino acid sequence of theformula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ IDNO: 189); ii) an amino acid sequence motif 4 as represented by an aminoacid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); iii) an amino acid sequence motif 7 as represented byan amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iv) an amino acid sequence motif 15 as representedby an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, at leastone amino acid sequence motif selected from: an amino acid sequencemotif 12 as represented by an amino acid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 as represented by anamino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 as represented by anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 17 as represented by anamino acid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif selected from: an amino acid sequencemotif 1 as represented by an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 as represented by an amino acidsequence of the formula [AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQID NO: 188); an amino acid sequence motif 3 as represented by an aminoacid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15 asrepresented by an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 as represented by an amino acid sequence ofthe formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 as represented by anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); and anamino acid sequence motif 4 as represented by an amino acid sequence ofthe formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); andat least one amino acid sequence motif selected from: i) an amino acidsequence motif 2 having at least 90% identity to an amino acid sequenceof the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); and ii) an amino acid sequence motif 3 having at least90% identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); and at least one amino acid sequencemotif selected from: i) an amino acid sequence motif 1 having at least90% identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); ii) an amino acid sequence motif 5 having at least 90% identity toan amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); iii) anamino acid sequence motif 12 having at least 90% identity to an aminoacid sequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); and iv) an amino acid sequence motif 16 having atleast 90% identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 11 having at least 90% identity to an amino acid sequenceof the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); ii) anamino acid sequence motif 17 having at least 90% identity to an aminoacid sequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); iii) an amino acid sequence motif 20 having at least 90% identityto an amino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV](SEQ ID NO:192); iv) an amino acid sequence motif 24 having at least 90% identityto an amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and v) an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 11 having at least 90% identity to an amino acidsequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 17 having at least 90% identity to an amino acidsequence of the formulaD[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE (SEQ ID NO:189); an amino acid sequence motif 20 having at least 90% identity to anamino acid sequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 24 having at least 90% identity to anamino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196); and an aminoacid sequence motif 26 having at least 90% identity to an amino acidsequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 11having at least 90% identity to an amino acid sequence of the formulaK[ST]F[NTQS]L[QGLNIV]N[TAS]A[DHNQE][QHN][DE]D[PQN]SST[EQND][QKNR]LAW[EQND]LADEP[SRTK]FDNV[SFTWY][HLIV][ST]W[SITLV]LSN (SEQ ID NO: 183); an aminoacid sequence motif 5 having at least 90% identity to an amino acidsequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 20 having at least 90% identity to an amino acidsequence of the formulaDT[QEND]SSSLSSSSSL[LSIVT]LPWSR[NSQT]GSLLPS[EKQDNR]S[LFIV] (SEQ ID NO:192); an amino acid sequence motif 26 having at least 90% identity to anamino acid sequence of the formulaL[SVTIL]AL[PTS][YFRWK][RDGQKEN]G[NDITQELVS][VALI][NSRQTK][GDNEQ]L[PFRSKT]F[NDETQS](SEQ ID NO: 197) and an amino acid sequence motif 24 having at least 90%identity to an amino acid sequence of the formula[AQN][RVKIL][VTILS]TT[ST][GH][GST][TRSK]FSF[SPT]GL[QHKNR]G[AFLRIVK][FY]TG[QEDN]S[HYW][VAIL][IPLV][ADE][IVL]T[TAS]Q (SEQ ID NO: 196).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 8 having at least 90% identity to an amino acid sequenceof the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); ii) an amino acid sequence motif 13 having at least90% identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); iii) an aminoacid sequence motif 19 having at least 90% identity to an amino acidsequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); iv) an amino acid sequence motif 22 having at least 90% identityto an amino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); and v)an amino acid sequence motif 23 having at least 90% identity to an aminoacid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 8 having at least 90% identity to an amino acidsequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE](SEQ ID NO: 185); an amino acidsequence motif 19 having at least 90% identity to an amino acid sequenceof the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 22 having at least 90% identity to anamino acid sequence of the formula[KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ ID NO: 194); anamino acid sequence motif 23 having at least 90% identity to an aminoacid sequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); andat least one amino acid sequence motif selected from: i) an amino acidsequence motif 4 having at least 90% identity to an amino acid sequenceof the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); ii) an amino acid sequence motif 7 having at least 90%identity to an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iii) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 having at least 90% identity to anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15having at least 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 having at least 90% identity to an aminoacid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 23 having at least 90% identity to an amino acidsequence of the formula Y[TAS][ST]TQEESRTLT (SEQ ID NO: 195); an aminoacid sequence motif 4 having at least 90% identity to an amino acidsequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); an amino acid sequence motif 13 having at least 90%identity to an amino acid sequence of the formulaGVDYTSVDI[TVIL][NSQT][TQSN]GTRSK[ADE] (SEQ ID NO: 185); an amino acidsequence motif 22 having at least 90% identity to an amino acid sequenceof the formula [KLSRIVT][DKNERQ][EHQND]VEV[EQDN]A[TVSIL][DE]QQ (SEQ IDNO: 194); and an amino acid sequence motif 8 having at least 90%identity to an amino acid sequence of the formula[QGN][EVD][GKR]D[HQN][NSQT]VQP[NDQE]KE[VAIL][EKQDRN]E[CRSKT][KTSR][LVI]LF[AITLVS]E(SEQ ID NO: 180).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide, an aminoacid sequence motif 2 having at least 90% identity to an amino acidsequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 19 having at least 90%identity to an amino acid sequence of the formula[TS]G[DSTNQE][DS][TSNQ]P[AHYRITKLVS][TVLSI][FYC]G[TS][APVIL] (SEQ ID NO:191); an amino acid sequence motif 6 having at least 90% identity to anamino acid sequence of the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT(SEQ ID NO: 178); an amino acid sequence motif 10 having at least 90%identity to an amino acid sequence of the formula[WPYF][DTNQES][HNVQIL][VIL][AG]R[VIL][RWKYH]F[SQTN]TS[RLKIV][GLIV]R[TISLV]FE[YFW]G[SMT][ST][RV][EQND]P[ST]G[QKRN][WVYIL][FITLVS][TESD][VAIL]NVGSGVCVG[VMIL] (SEQ ID NO: 182); an amino acid sequence motif 3 having at least90% identity to an amino acid sequence ofGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 27having at least 90% identity to an amino acid sequence of the formulaT[TFPS]IL[EQSDNT][EQHDN]LP[DAE] (SEQ ID NO: 198); an amino acid sequencemotif 7 having at least 90% identity to an amino acid sequence of theformula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 21 having at least 90%identity to an amino acid sequence of the formulaHL[TVSIL][GA][NRSQKT][ANDQE]I[STLIV][IVL][NDQE][VILM][DTSE]LGI[ND][TAS]P[TKSR][VIL]V[PA](SEQ ID NO: 193); an amino acid sequence motif 18 having at least 90%identity to an amino acid sequence of the formulaG[TLPSIV][AED]GAST[NTAQS]F[GQHN]W[EQND]T[VAIL]R[TAS]FPS[TISLV]NA (SEQ IDNO: 190); an amino acid sequence motif 9 having at least 90% identity toan amino acid sequence of the formula[IMVL][QEND]G[ST]I[AST][NDQE]L[TISLV]V[SRTK]T[NADQE][EAD]YSVW[CG]H[IVL]SDT[LVI]APAQ[SLTIV][LIV][PIVL][KSRT][HRK][RASVKTIL]A (SEQ ID NO: 181); anamino acid sequence motif 14 having at least 90% identity to an aminoacid sequence of the formula[LWIVF][VIL]GEG[RSKT]I[TDSE][ANQ]L[PQN]CSA[NRQK]IQV[FILV]T[SDTE][GAST][GADE][NGYQ][NDGQE][LFIV]PF[AG][TS]F[ST]FPV[RGQKN][LSVIT][LFIV]Y[DNSQET][GA][GRK]AHS[DNQTES]VQ[VIL][LIV] (SEQ ID NO: 186).

In some embodiments a PtIP-65 polypeptide comprises an amino acidsequence motif 6 having at least 90% identity to an amino acid sequenceof the formula R[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); anamino acid sequence motif 2 having at least 90% identity to an aminoacid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 3 having at least 90%identity to an amino acid sequence ofGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motifselected from: an amino acid sequence motif 1 having at least 90%identity to an amino acid sequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 5 having at least 90% identity to anamino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 16 having at least 90%identity to an amino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); and atleast one amino acid sequence motif selected from: i) an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); ii) an amino acid sequence motif 4 having at least 90%identity to an amino acid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176); iii) an amino acid sequence motif 7 having at least90% identity to an amino acid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); and iv) an amino acid sequence motif 15 having atleast 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187).

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus of the PtIP-65 polypeptide: an aminoacid sequence motif 12 having at least 90% identity to an amino acidsequence of the formula[LMIV][AVYILF][QLNIV][TVSIL][PILV][VGIL][YTWS][VLI]IGGQGG[SNTQ][AQSNT](SEQ ID NO: 184); an amino acid sequence motif 2 having at least 90%identity to an amino acid sequence of the formulaF[STRK]Y[DYEWF][GQN][GSETD][TRKS][DNQE]G[RK]V[LMIV][QTRNSK][RK]I[GRK]V[WY][ARK][GA][ETDS][SWTYF][RQKN][ILV][KR][AG]I[RESKDT]VW[LMIV](SEQ ID NO: 174); an amino acid sequence motif 6 having at least 90%identity to an amino acid sequence of the formulaR[GLIV][SETD]YSE[FYW]TFA[DETS]GERIT (SEQ ID NO: 178); an amino acidsequence motif 17 having at least 90% identity to an amino acid sequenceof the formula D[GED][VAIL][QEAND][KQRN][TLSIV]FGDP[SHT][RDKE]P[APST]GE(SEQ ID NO: 189); an amino acid sequence motif selected from: an aminoacid sequence motif 1 having at least 90% identity to an amino acidsequence of the formulaRLSLWGNGAGTR[SAT]G[WAGYF]I[RYKWF]F[YERDKWF]T[TNSQ][TQNS][GNQ][GRK][SRTK]F[FDE][FPH][GKR]M[TYSWF]SWG[LKRIV][KQRN]TEYP[IVQLN]DV (SEQ ID NO:173); an amino acid sequence motif 16 having at least 90% identity to anamino acid sequence of the formula[AVSILT]SG[ILV][CWY][VAIL]G[IVLF][TILMRSVK] (SEQ ID NO: 188); an aminoacid sequence motif 3 having at least 90% identity to an amino acidsequence of the formulaGRA[NSQT][SVTIL]D[IVL][DNQE][SAT]LG[FVIL][VMLI]FLR[PTS]I[QAN]S[AVCSILT]R[LMIV]I[NSQT]V[QSNT]YPTL (SEQ ID NO: 175); an amino acid sequence motif 15having at least 90% identity to an amino acid sequence of the formula[LFIV][ED][QTNS][AQ]GI[IVSTL]P[VITQLSN][TAS]L[DKER] (SEQ ID NO: 187); anamino acid sequence motif 7 having at least 90% identity to an aminoacid sequence of the formula[NTQS]D[SKTR]NN[ADE][GDSET]S[IELVD][PST][KLRIV]NW[TVSIL][FLIV][SAT]GSR[TQSN]VT[ITVLS]SS[ST]W[STRK][LTIVS]T(SEQ ID NO: 179); an amino acid sequence motif 5 having at least 90%identity to an amino acid sequence of the formula[GQN][ILFV][EITDLVS][ATVSIL][HTS][AST]S[VIL][TS]V[QESNDT]A[GRK]IP[AST][VLI]AEV[SNTQ]G[EVDIL][FVIL]GW[SEQTDN][VIL]S[VAIL][ST] (SEQ ID NO: 177); and anamino acid sequence motif 4 having at least 90% identity to an aminoacid sequence of the formulaW[NGQ][QVNIL]SGTL[ESDT][PA]G[QEND][WSYT]I[SHT][LIV][QKNR]A[TLSIV]TR[RK]G[TLSIV]I[TS][LVI]P[YFW]Q[GA][TS][MILV][EQND][IVL]TL[QKLNRIV][SNTQ]G[TDSE][VIL]F[QRNK]Y[PA][ILV][SKTR][SGT][MQN]Y(SEQ ID NO: 176).

In some embodiments a PtIP-65 polypeptide is about 300 to 400 aminoacids in length having a consensus secondary structure comprising 13 to17 segments of predominately beta strand structure separated by coilregions.

In some embodiments a PtIP-65 polypeptide comprises, sequentially fromthe N-terminus to the C-terminus; i) a beta strand-1 (β1) of betweenabout 3 amino acids and about 9 amino acids in length, ii) a coil ofbetween about 17 amino acids and about 21 amino acids in length; iii) abeta strand-2 (β2) of between about 7 amino acids and about 9 aminoacids in length, iv) a coil of between about 3 amino acids and about 4amino acids in length; v) a beta strand-3 (β3) of between about 9 aminoacids and about 10 amino acids in length, vi) a coil of between about 2amino acids and about 5 amino acids in length; vii) a beta strand-4 (β4)of between about 3 amino acids and about 8 amino acids in length, viii)a coil of between about 7 amino acids and about 13 amino acids inlength; ix) a beta strand-5 (β5) of between about 3 amino acids andabout 5 amino acids in length, x) a coil of between about 4 amino acidsand about 5 amino acids in length; xi) a beta strand-6 (β6) of betweenabout 8 amino acids and about 12 amino acids in length, xii) a coil ofbetween about 5 amino acids and about 12 amino acids in length; xiii) abeta strand-7 (β7) of between about 9 amino acids and about 13 aminoacids in length, xiv) a coil of about 4 amino acids in length; xv) abeta strand-8 (β1) of between about 3 amino acids and about 7 aminoacids in length, xvi) a coil of between about 5 amino acids and about 9amino acids in length; xvii) a beta strand-9 (β9) of between about 3amino acids and about 5 amino acids in length, xviii) a coil of betweenabout 3 amino acids and about 6 amino acids in length; ixx) a betastrand-10 (β10) of between about 7 amino acids and about 13 amino acidsin length, xx) a coil of between about 3 amino acids and about 8 aminoacids in length; xxi) a beta strand-11 (β11) of between about 5 aminoacids and about 18 amino acids in length, xxii) a coil of between about10 amino acids and about 22 amino acids in length; xxiii) a betastrand-12 (β12) of between about 7 amino acids and about 12 amino acidsin length, xxiv) a coil of between about 5 amino acids and about 13amino acids in length; xxv) a beta strand-13 (β13) of between about 60amino acids and about 90 amino acids in length, xxvi) a coil of betweenabout 4 amino acids and about 8 amino acids in length; xxvii) a betastrand-14 (β14) of between about 22 amino acids and about 30 amino acidsin length, xxviii) a coil of between about 3 amino acids and about 8amino acids in length; and xxix) a beta strand-15 (β15) of between about22 amino acids and about 26 amino acids in length. As used herein, theterm “about” when used in the context of the lower/upper limit of thelength of a secondary structural element means the greater of −/+aninteger of up to −/+20% of the length of the secondary structuralelement or −/+1 amino acid. By means of example, a secondary structureelement of between about 3 amino acids and about 23 amino acids inlength means a secondary structure element of between 2 and 27 aminoacids in length.

In some embodiments a PtIP-65 polypeptide has a calculated molecularweight of between about 25kD and about 50kD, between about 27.5kD andabout 47.5kD, or between about 30kD and about 45kD.

In some embodiments the PtIP-65 polypeptide has a modified physicalproperty. As used herein, the term “physical property” refers to anyparameter suitable for describing the physical-chemical characteristicsof a protein. As used herein, “physical property of interest” and“property of interest” are used interchangeably to refer to physicalproperties of proteins that are being investigated and/or modified.Examples of physical properties include, but are not limited to netsurface charge and charge distribution on the protein surface, nethydrophobicity and hydrophobic residue distribution on the proteinsurface, surface charge density, surface hydrophobicity density, totalcount of surface ionizable groups, surface tension, protein size and itsdistribution in solution, melting temperature, heat capacity, and secondvirial coefficient. Examples of physical properties also include, butare not limited to solubility, folding, stability, and digestibility. Insome embodiments the PtIP-65 polypeptide has increased digestibility ofproteolytic fragments in an insect gut. Models for digestion bysimulated gastric fluids are known to one skilled in the art (Fuchs, R.L. and J. D. Astwood. Food Technology 50: 83-88, 1996; Astwood, J. D.,et al Nature Biotechnology 14: 1269-1273, 1996; Fu T J et al J. AgricFood Chem. 50: 7154-7160, 2002).

In some embodiments variants include polypeptides that differ in aminoacid sequence due to mutagenesis. Variant proteins encompassed by thedisclosure are biologically active, that is they continue to possess thedesired biological activity (i.e. pesticidal activity) of the nativeprotein. In some embodiment the variant will have at least about 10%, atleast about 30%, at least about 50%, at least about 70%, at least about80% or more of the insecticidal activity of the native protein. In someembodiments, the variants may have improved activity over the nativeprotein.

Bacterial genes quite often possess multiple methionine initiationcodons in proximity to the start of the open reading frame. Often,translation initiation at one or more of these start codons will lead togeneration of a functional protein. These start codons can include ATGcodons. However, bacteria such as Bacillus sp. also recognize the codonGTG as a start codon, and proteins that initiate translation at GTGcodons contain a methionine at the first amino acid. On rare occasions,translation in bacterial systems can initiate at a TTG codon, though inthis event the TTG encodes a methionine. Furthermore, it is not oftendetermined a priori which of these codons are used naturally in thebacterium. Thus, it is understood that use of one of the alternatemethionine codons may also lead to generation of pesticidal proteins.These pesticidal proteins are encompassed in the present disclosure andmay be used in the methods of the present disclosure. It will beunderstood that, when expressed in plants, it will be necessary to alterthe alternate start codon to ATG for proper translation.

In another aspect the PtIP-50 polypeptide or PtIP-65 polypeptide may beexpressed as a precursor protein with an intervening sequence thatcatalyzes multi-step, post translational protein splicing. Proteinsplicing involves the excision of an intervening sequence from apolypeptide with the concomitant joining of the flanking sequences toyield a new polypeptide (Chong, et al., (1996) J. Biol. Chem.,271:22159-22168). This intervening sequence or protein splicing element,referred to as inteins, which catalyze their own excision through threecoordinated reactions at the N-terminal and C-terminal splice junctions:an acyl rearrangement of the N-terminal cysteine or serine; atransesterification reaction between the two termini to form a branchedester or thioester intermediate and peptide bond cleavage coupled tocyclization of the intein C-terminal asparagine to free the intein(Evans, et al., (2000) J. Biol. Chem., 275:9091-9094. The elucidation ofthe mechanism of protein splicing has led to a number of intein-basedapplications (Comb, et al., U.S. Pat. No. 5,496,714; Comb, et al., U.S.Pat. No. 5,834,247; Camarero and Muir, (1999) J. Amer. Chem. Soc.121:5597-5598; Chong, et al., (1997) Gene 192:271-281, Chong, et al.,(1998) Nucleic Acids Res. 26:5109-5115; Chong, et al., (1998) J. Biol.Chem. 273:10567-10577; Cotton, et al., (1999) J. Am. Chem. Soc.121:1100-1101; Evans, et al., (1999) J. Biol. Chem. 274:18359-18363;Evans, et al., (1999) J. Biol. Chem. 274:3923-3926; Evans, et al.,(1998) Protein Sci. 7:2256-2264; Evans, et al., (2000) J. Biol. Chem.275:9091-9094; Iwai and Pluckthun, (1999) FEBS Lett. 459:166-172;Mathys, et al., (1999) Gene 231:1-13; Mills, et al., (1998) Proc. Natl.Acad. Sci. USA 95:3543-3548; Muir, et al., (1998) Proc. Natl. Acad. Sci.USA 95:6705-6710; Otomo, et al., (1999) Biochemistry 38:16040-16044;Otomo, et al., (1999) J. Biolmol. NMR 14:105-114; Scott, et al., (1999)Proc. Natl. Acad. Sci. USA 96:13638-13643; Severinov and Muir, (1998) J.Biol. Chem. 273:16205-16209; Shingledecker, et al., (1998) Gene207:187-195; Southworth, et al., (1998) EMBO J. 17:918-926; Southworth,et al., (1999) Biotechniques 27:110-120; Wood, et al., (1999) Nat.Biotechnol. 17:889-892; Wu, et al., (1998a) Proc. Natl. Acad. Sci. USA95:9226-9231; Wu, et al., (1998b) Biochim Biophys Acta 1387:422-432; Xu,et al., (1999) Proc. Natl. Acad. Sci. USA 96:388-393; Yamazaki, et al.,(1998) J. Am. Chem. Soc., 120:5591-5592). For the application of inteinsin plant transgenes, see, Yang, et al., (Transgene Res 15:583-593(2006)) and Evans, et al., (Annu. Rev. Plant Biol. 56:375-392 (2005)).

In another aspect the PtIP-50 polypeptide or PtIP-65 polypeptide may beencoded by two separate genes where the intein of the precursor proteincomes from the two genes, referred to as a split-intein, and the twoportions of the precursor are joined by a peptide bond formation. Thispeptide bond formation is accomplished by intein-mediatedtrans-splicing. For this purpose, a first and a second expressioncassette comprising the two separate genes further code for inteinscapable of mediating protein trans-splicing. By trans-splicing, theproteins and polypeptides encoded by the first and second fragments maybe linked by peptide bond formation. Trans-splicing inteins may beselected from the nucleolar and organellar genomes of differentorganisms including eukaryotes, archaebacteria and eubacteria. Inteinsthat may be used for are listed at neb.com/neb/inteins.html, which canbe accessed on the world-wide web using the “www” prefix). Thenucleotide sequence coding for an intein may be split into a 5′ and a 3′part that code for the 5′ and the 3′ part of the intein, respectively.Sequence portions not necessary for intein splicing (e.g. homingendonuclease domain) may be deleted. The intein coding sequence is splitsuch that the 5′ and the 3′ parts are capable of trans-splicing. Forselecting a suitable splitting site of the intein coding sequence, theconsiderations published by Southworth, et al., (1998) EMBO J.17:918-926 may be followed. In constructing the first and the secondexpression cassette, the 5′ intein coding sequence is linked to the 3′end of the first fragment coding for the N-terminal part of the PtIP-50polypeptide or PtIP-65 polypeptide and the 3′ intein coding sequence islinked to the 5′ end of the second fragment coding for the C-terminalpart of the PtIP-50 polypeptide or PtIP-65 polypeptide.

In general, the trans-splicing partners can be designed using any splitintein, including any naturally-occurring or artificially-split splitintein. Several naturally-occurring split inteins are known, forexample: the split intein of the DnaE gene of Synechocystis sp. PCC6803(see, Wu, et al., (1998) Proc Natl Acad Sci USA. 95(16):9226-31 andEvans, et al., (2000) J Biol Chem. 275(13):9091-4 and of the DnaE genefrom Nostoc punctiforme (see, Iwai, et al., (2006) FEBS Lett.580(7):1853-8). Non-split inteins have been artificially split in thelaboratory to create new split inteins, for example: the artificiallysplit Ssp DnaB intein (see, Wu, et al., (1998) Biochim Biophys Acta.1387:422-32) and split Sce VMA intein (see, Brenzel, et al., (2006)Biochemistry. 45(6):1571-8) and an artificially split fungal mini-intein(see, Elleuche, et al., (2007) Biochem Biophys Res Commun.355(3):830-4). There are also intein databases available that catalogueknown inteins (see for example the online-database available at:bioinformatics.weizmann.ac.ilrpietro/inteins/Inteinstable.html, whichcan be accessed on the world-wide web using the “www” prefix).

Naturally-occurring non-split inteins may have endonuclease or otherenzymatic activities that can typically be removed when designing anartificially-split split intein. Such mini-inteins or minimized splitinteins are well known in the art and are typically less than 200 aminoacid residues long (see, Wu, et al., (1998) Biochim Biophys Acta.1387:422-32). Suitable split inteins may have other purificationenabling polypeptide elements added to their structure, provided thatsuch elements do not inhibit the splicing of the split intein or areadded in a manner that allows them to be removed prior to splicing.Protein splicing has been reported using proteins that comprisebacterial intein-like (BIL) domains (see, Amitai, et al., (2003) MolMicrobiol. 47:61-73) and hedgehog (Hog) auto-processing domains (thelatter is combined with inteins when referred to as the Hog/inteinsuperfamily or HINT family (see, Dassa, et al., (2004) J Biol Chem.279:32001-7) and domains such as these may also be used to prepareartificially-split inteins. In particular, non-splicing members of suchfamilies may be modified by molecular biology methodologies to introduceor restore splicing activity in such related species. Recent studiesdemonstrate that splicing can be observed when a N-terminal split inteincomponent is allowed to react with a C-terminal split intein componentnot found in nature to be its “partner”; for example, splicing has beenobserved utilizing partners that have as little as 30 to 50% homologywith the “natural” splicing partner (see, Dassa, et al., (2007)Biochemistry. 46(1):322-30). Other such mixtures of disparate splitintein partners have been shown to be unreactive one with another (see,Brenzel, et al., (2006) Biochemistry. 45(6):1571-8). However, it iswithin the ability of a person skilled in the relevant art to determinewhether a particular pair of polypeptides is able to associate with eachother to provide a functional intein, using routine methods and withoutthe exercise of inventive skill.

In another aspect the PtIP-50 polypeptide is a circular permutedvariant. In certain embodiments the PtIP-50 polypeptide is a circularpermuted variant of the polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO:54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ IDNO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68,SEQ ID NO: 69 or SEQ ID NO: 70.

In another aspect the PtIP-65 polypeptide is a circular permutedvariant. In certain embodiments the PtIP-65 polypeptide is a circularpermuted variant of the polypeptide of SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO:33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ IDNO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42.

The development of recombinant DNA methods has made it possible to studythe effects of sequence transposition on protein folding, structure andfunction. The approach used in creating new sequences resembles that ofnaturally occurring pairs of proteins that are related by linearreorganization of their amino acid sequences (Cunningham, et al., (1979)Proc. Natl. Acad. Sci. U.S.A. 76:3218-3222; Teather and Erfle, (1990) J.Bacteriol. 172:3837-3841; Schimming, et al., (1992) Eur. J. Biochem.204:13-19; Yamiuchi and Minamikawa, (1991) FEBS Lett. 260:127-130;MacGregor, et al., (1996) FEBS Lett. 378:263-266). The first in vitroapplication of this type of rearrangement to proteins was described byGoldenberg and Creighton (J. Mol. Biol. 165:407-413, 1983). In creatinga circular permuted variant a new N-terminus is selected at an internalsite (breakpoint) of the original sequence, the new sequence having thesame order of amino acids as the original from the breakpoint until itreaches an amino acid that is at or near the original C-terminus. Atthis point the new sequence is joined, either directly or through anadditional portion of sequence (linker), to an amino acid that is at ornear the original N-terminus and the new sequence continues with thesame sequence as the original until it reaches a point that is at ornear the amino acid that was N-terminal to the breakpoint site of theoriginal sequence, this residue forming the new C-terminus of the chain.The length of the amino acid sequence of the linker can be selectedempirically or with guidance from structural information or by using acombination of the two approaches. When no structural information isavailable, a small series of linkers can be prepared for testing using adesign whose length is varied in order to span a range from 0 to 50 Åand whose sequence is chosen in order to be consistent with surfaceexposure (hydrophilicity, Hopp and Woods, (1983) Mol. Immunol.20:483-489; Kyte and Doolittle, (1982) J. Mol. Biol. 157:105-132;solvent exposed surface area, Lee and Richards, (1971) J. Mol. Biol.55:379-400) and the ability to adopt the necessary conformation withoutderanging the configuration of the pesticidal polypeptide(conformationally flexible; Karplus and Schulz, (1985)Naturwissenschaften 72:212-213). Assuming an average of translation of2.0 to 3.8 Å per residue, this would mean the length to test would bebetween 0 to 30 residues, with 0 to 15 residues being the preferredrange. Exemplary of such an empirical series would be to constructlinkers using a cassette sequence such as Gly-Gly-Gly-Ser repeated ntimes, where n is 1, 2, 3 or 4. Those skilled in the art will recognizethat there are many such sequences that vary in length or compositionthat can serve as linkers with the primary consideration being that theybe neither excessively long nor short (cf., Sandhu, (1992) Critical Rev.Biotech. 12:437-462); if they are too long, entropy effects will likelydestabilize the three-dimensional fold, and may also make foldingkinetically impractical, and if they are too short, they will likelydestabilize the molecule because of torsional or steric strain. Thoseskilled in the analysis of protein structural information will recognizethat using the distance between the chain ends, defined as the distancebetween the c-alpha carbons, can be used to define the length of thesequence to be used or at least to limit the number of possibilitiesthat must be tested in an empirical selection of linkers. They will alsorecognize that it is sometimes the case that the positions of the endsof the polypeptide chain are ill-defined in structural models derivedfrom x-ray diffraction or nuclear magnetic resonance spectroscopy data,and that when true, this situation will therefore need to be taken intoaccount in order to properly estimate the length of the linker required.From those residues whose positions are well defined are selected tworesidues that are close in sequence to the chain ends, and the distancebetween their c-alpha carbons is used to calculate an approximate lengthfor a linker between them. Using the calculated length as a guide,linkers with a range of number of residues (calculated using 2 to 3.8 Åper residue) are then selected. These linkers may be composed of theoriginal sequence, shortened or lengthened as necessary, and whenlengthened the additional residues may be chosen to be flexible andhydrophilic as described above; or optionally the original sequence maybe substituted for using a series of linkers, one example being theGly-Gly-Gly-Ser cassette approach mentioned above; or optionally acombination of the original sequence and new sequence having theappropriate total length may be used. Sequences of pesticidalpolypeptides capable of folding to biologically active states can beprepared by appropriate selection of the beginning (amino terminus) andending (carboxyl terminus) positions from within the originalpolypeptide chain while using the linker sequence as described above.Amino and carboxyl termini are selected from within a common stretch ofsequence, referred to as a breakpoint region, using the guidelinesdescribed below. A novel amino acid sequence is thus generated byselecting amino and carboxyl termini from within the same breakpointregion. In many cases the selection of the new termini will be such thatthe original position of the carboxyl terminus immediately preceded thatof the amino terminus. However, those skilled in the art will recognizethat selections of termini anywhere within the region may function, andthat these will effectively lead to either deletions or additions to theamino or carboxyl portions of the new sequence. It is a central tenet ofmolecular biology that the primary amino acid sequence of a proteindictates folding to the three-dimensional structure necessary forexpression of its biological function. Methods are known to thoseskilled in the art to obtain and interpret three-dimensional structuralinformation using x-ray diffraction of single protein Crystals ornuclear magnetic resonance spectroscopy of protein solutions. Examplesof structural information that are relevant to the identification ofbreakpoint regions include the location and type of protein secondarystructure (alpha and 3-10 helices, parallel and anti-parallel betasheets, chain reversals and turns, and loops; Kabsch and Sander, (1983)Biopolymers 22:2577-2637; the degree of solvent exposure of amino acidresidues, the extent and type of interactions of residues with oneanother (Chothia, (1984) Ann. Rev. Biochem. 53:537-572) and the staticand dynamic distribution of conformations along the polypeptide chain(Alber and Mathews, (1987) Methods Enzymol. 154:511-533). In some casesadditional information is known about solvent exposure of residues; oneexample is a site of post-translational attachment of carbohydrate whichis necessarily on the surface of the protein. When experimentalstructural information is not available or is not feasible to obtain,methods are also available to analyze the primary amino acid sequence inorder to make predictions of protein tertiary and secondary structure,solvent accessibility and the occurrence of turns and loops. Biochemicalmethods are also sometimes applicable for empirically determiningsurface exposure when direct structural methods are not feasible; forexample, using the identification of sites of chain scission followinglimited proteolysis in order to infer surface exposure (Gentile andSalvatore, (1993) Eur. J. Biochem. 218:603-621). Thus using either theexperimentally derived structural information or predictive methods(e.g., Srinivisan and Rose, (1995) Proteins: Struct., Funct. & Genetics22:81-99) the parental amino acid sequence is inspected to classifyregions according to whether or not they are integral to the maintenanceof secondary and tertiary structure. The occurrence of sequences withinregions that are known to be involved in periodic secondary structure(alpha and 3-10 helices, parallel and anti-parallel beta sheets) areregions that should be avoided. Similarly, regions of amino acidsequence that are observed or PtIP- to have a low degree of solventexposure are more likely to be part of the so-called hydrophobic core ofthe protein and should also be avoided for selection of amino andcarboxyl termini. In contrast, those regions that are known or PtIP- tobe in surface turns or loops, and especially those regions that areknown not to be required for biological activity, are the preferredsites for location of the extremes of the polypeptide chain. Continuousstretches of amino acid sequence that are preferred based on the abovecriteria are referred to as a breakpoint region. Polynucleotidesencoding circular permuted PtIP-50 polypeptides or PtIP-65 polypeptideswith new N-terminus/C-terminus which contain a linker region separatingthe original C-terminus and N-terminus can be made essentially followingthe method described in Mullins, et al., (1994) J. Am. Chem. Soc.116:5529-5533. Multiple steps of polymerase chain reaction (PCR)amplifications are used to rearrange the DNA sequence encoding theprimary amino acid sequence of the protein. Polynucleotides encodingcircular permuted PtIP-50 polypeptides or PtIP-65 polypeptides with newN-terminus/C-terminus which contain a linker region separating theoriginal C-terminus and N-terminus can be made based on thetandem-duplication method described in Horlick, et al., (1992) ProteinEng. 5:427-431. Polymerase chain reaction (PCR) amplification of the newN-terminus/C-terminus genes is performed using a tandemly duplicatedtemplate DNA.

In another aspect fusion proteins are provided that include within itsamino acid sequence an amino acid sequence comprising a PtIP-50polypeptide including but not limited to the polypeptide of SEQ ID NO:43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ IDNO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57,SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO:62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ IDNO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ ID NO: 70, and activefragments thereof.

In another aspect fusion proteins are provided that include within itsamino acid sequence an amino acid sequence comprising a PtIP-65polypeptide including but not limited to the polypeptide of SEQ ID NO:22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ IDNO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36,SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO:41 or SEQ ID NO: 42, and active fragments thereof.

Methods for design and construction of fusion proteins (andpolynucleotides encoding same) are known to those of skill in the art.Polynucleotides encoding a PtIP-50 polypeptide or PtIP-65 polypeptidemay be fused to signal sequences which will direct the localization ofthe PtIP-50 polypeptide or PtIP-65 polypeptide to particularcompartments of a prokaryotic or eukaryotic cell and/or direct thesecretion of the PtIP-50 polypeptide or PtIP-65 polypeptide of theembodiments from a prokaryotic or eukaryotic cell. For example, in E.coli, one may wish to direct the expression of the protein to theperiplasmic space. Examples of signal sequences or proteins (orfragments thereof) to which the PtIP-50 polypeptide or PtIP-65polypeptide may be fused in order to direct the expression of thepolypeptide to the periplasmic space of bacteria include, but are notlimited to, the pelB signal sequence, the maltose binding protein (MBP)signal sequence, MBP, the ompA signal sequence, the signal sequence ofthe periplasmic E. coli heat-labile enterotoxin B-subunit and the signalsequence of alkaline phosphatase. Several vectors are commerciallyavailable for the construction of fusion proteins which will direct thelocalization of a protein, such as the pMAL series of vectors(particularly the pMAL-p series) available from New England Biolabs. Ina specific embodiment, the PtIP-50 polypeptide or PtIP-65 polypeptidemay be fused to the pelB pectate lyase signal sequence to increase theefficiency of expression and purification of such polypeptides inGram-negative bacteria (see, U.S. Pat. Nos. 5,576,195 and 5,846,818).Plant plastid transit peptide/polypeptide fusions are well known in theart (see, U.S. Pat. No. 7,193,133). Apoplast transit peptides such asrice or barley alpha-amylase secretion signal are also well known in theart. The plastid transit peptide is generally fused N-terminal to thepolypeptide to be targeted (e.g., the fusion partner). In oneembodiment, the fusion protein consists essentially of the plastidtransit peptide and the PtIP-50 polypeptide or PtIP-65 polypeptide to betargeted. In another embodiment, the fusion protein comprises theplastid transit peptide and the polypeptide to be targeted. In suchembodiments, the plastid transit peptide is preferably at the N-terminusof the fusion protein. However, additional amino acid residues may beN-terminal to the plastid transit peptide providing that the fusionprotein is at least partially targeted to a plastid. In a specificembodiment, the plastid transit peptide is in the N-terminal half,N-terminal third or N-terminal quarter of the fusion protein. Most orall of the plastid transit peptide is generally cleaved from the fusionprotein upon insertion into the plastid. The position of cleavage mayvary slightly between plant species, at different plant developmentalstages, as a result of specific intercellular conditions or theparticular combination of transit peptide/fusion partner used. In oneembodiment, the plastid transit peptide cleavage is homogenous such thatthe cleavage site is identical in a population of fusion proteins. Inanother embodiment, the plastid transit peptide is not homogenous, suchthat the cleavage site varies by 1-10 amino acids in a population offusion proteins. The plastid transit peptide can be recombinantly fusedto a second protein in one of several ways. For example, a restrictionendonuclease recognition site can be introduced into the nucleotidesequence of the transit peptide at a position corresponding to itsC-terminal end and the same or a compatible site can be engineered intothe nucleotide sequence of the protein to be targeted at its N-terminalend. Care must be taken in designing these sites to ensure that thecoding sequences of the transit peptide and the second protein are kept“in frame” to allow the synthesis of the desired fusion protein. In somecases, it may be preferable to remove the initiator methionine codon ofthe second protein when the new restriction site is introduced. Theintroduction of restriction endonuclease recognition sites on bothparent molecules and their subsequent joining through recombinant DNAtechniques may result in the addition of one or more extra amino acidsbetween the transit peptide and the second protein. This generally doesnot affect targeting activity as long as the transit peptide cleavagesite remains accessible and the function of the second protein is notaltered by the addition of these extra amino acids at its N-terminus.Alternatively, one skilled in the art can create a precise cleavage sitebetween the transit peptide and the second protein (with or without itsinitiator methionine) using gene synthesis (Stemmer, et al., (1995) Gene164:49-53) or similar methods. In addition, the transit peptide fusioncan intentionally include amino acids downstream of the cleavage site.The amino acids at the N-terminus of the mature protein can affect theability of the transit peptide to target proteins to plastids and/or theefficiency of cleavage following protein import. This may be dependenton the protein to be targeted. See, e.g., Comai, et al., (1988) J. Biol.Chem. 263(29):15104-9.

In some embodiments fusion proteins are provide comprising a PtIP-50polypeptide or PtIP-65 polypeptide and an insecticidal polypeptidejoined by an amino acid linker. In some embodiments fusion proteins areprovide comprising a PtIP-50 polypeptide and a PtIP-65 polypeptidejoined by an amino acid linker.

In some embodiments fusion proteins are provided represented by aformula selected from the group consisting of:R¹-L-R², R²-L-R¹, R¹-R² or R²-R¹

wherein R¹ is a PtIP-50 polypeptide, R² is PtIP-65 polypeptide. The R¹polypeptide is fused either directly or through a linker (L) segment tothe R² polypeptide. The term “directly” defines fusions in which thepolypeptides are joined without a peptide linker. Thus “L” represents achemical bound or polypeptide segment to which both R¹ and R² are fusedin frame, most commonly L is a linear peptide to which R¹ and R² arebound by amide bonds linking the carboxy terminus of R¹ to the aminoterminus of L and carboxy terminus of L to the amino terminus of R². By“fused in frame” is meant that there is no translation termination ordisruption between the reading frames of R¹ and R². The linking group(L) is generally a polypeptide of between 1 and 500 amino acids inlength. The linkers joining the two molecules are preferably designed to(1) allow the two molecules to fold and act independently of each other,(2) not have a propensity for developing an ordered secondary structurewhich could interfere with the functional domains of the two proteins,(3) have minimal hydrophobic or charged characteristic which couldinteract with the functional protein domains and (4) provide stericseparation of R¹ and R² such that R¹ and R² could interactsimultaneously with their corresponding receptors on a single cell.Typically surface amino acids in flexible protein regions include Gly,Asn and Ser. Virtually any permutation of amino acid sequencescontaining Gly, Asn and Ser would be expected to satisfy the abovecriteria for a linker sequence. Other neutral amino acids, such as Thrand Ala, may also be used in the linker sequence. Additional amino acidsmay also be included in the linkers due to the addition of uniquerestriction sites in the linker sequence to facilitate construction ofthe fusions.

In some embodiments the linkers comprise sequences selected from thegroup of formulas: (Gly₃Ser)_(n), (Gly₄Ser)_(n), (Gly₅Ser)_(n),(Gly_(n)Ser)_(n) or (AlaGlySer)_(n) where n is an integer. One exampleof a highly-flexible linker is the (GlySer)-rich spacer region presentwithin the pill protein of the filamentous bacteriophages, e.g.bacteriophages M13 or fd (Schaller, et al., 1975). This region providesa long, flexible spacer region between two domains of the pill surfaceprotein. Also included are linkers in which an endopeptidase recognitionsequence is included. Such a cleavage site may be valuable to separatethe individual components of the fusion to determine if they areproperly folded and active in vitro. Examples of various endopeptidasesinclude, but are not limited to, Plasmin, Enterokinase, Kallikerin,Urokinase, Tissue Plasminogen activator, clostripain, Chymosin,Collagenase, Russell's Viper Venom Protease, Postproline cleavageenzyme, V8 protease, Thrombin and factor Xa. In some embodiments thelinker comprises the amino acids EEKKN (SEQ ID NO: 201) from themulti-gene expression vehicle (MGEV), which is cleaved by vacuolarproteases as disclosed in US Patent Application Publication Number US2007/0277263. In other embodiments, peptide linker segments from thehinge region of heavy chain immunoglobulins IgG, IgA, IgM, IgD or IgEprovide an angular relationship between the attached polypeptides.Especially useful are those hinge regions where the cysteines arereplaced with serines. Linkers of the present disclosure includesequences derived from murine IgG gamma 2b hinge region in which thecysteines have been changed to serines. The fusion proteins are notlimited by the form, size or number of linker sequences employed and theonly requirement of the linker is that functionally it does notinterfere adversely with the folding and function of the individualmolecules of the fusion.

In another aspect chimeric PtIP-50 polypeptides are provided that arecreated through joining two or more portions of PtIP-50 genes, whichoriginally encoded separate PtIP-50 proteins to create a chimeric gene.The translation of the chimeric gene results in a single chimericPtIP-50 polypeptide with regions, motifs or domains derived from each ofthe original polypeptides. In certain embodiments the chimeric proteincomprises portions, motifs or domains of PtIP-50 polypeptides of SEQ IDNO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52,SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ IDNO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ ID NO: 70 in anycombination.

In another aspect chimeric PtIP-65 polypeptides are provided that arecreated through joining two or more portions of PtIP-65 genes, whichoriginally encoded separate PtIP-65 proteins to create a chimeric gene.The translation of the chimeric gene results in a single chimericPtIP-65 polypeptide with regions, motifs or domains derived from each ofthe original polypeptides. In certain embodiments the chimeric proteincomprises portions, motifs or domains of the PtIP-65 polypeptides of SEQID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26,SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ IDNO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQID NO: 41 or SEQ ID NO: 42 in any combination.

It is recognized that DNA sequences may be altered by various methods,and that these alterations may result in DNA sequences encoding proteinswith amino acid sequences different than that encoded by the wild-type(or native) pesticidal protein. In some embodiments a PtIP-50polypeptide or PtIP-65 polypeptide may be altered in various waysincluding amino acid substitutions, deletions, truncations andinsertions of one or more amino acids, including up to 2, 3, 4, 5, 6, 7,8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145 or more amino acidsubstitutions, deletions and/or insertions or combinations thereofcompared to any one of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ IDNO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69,SEQ ID NO: 70, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ IDNO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39,SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42.

In some embodiments a PtIP-50 polypeptide or PtIP-65 polypeptidecomprises a deletion of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15 or more amino acids from the N-terminus of the PtIP-50 polypeptiderelative to the amino acid position of any one of SEQ ID NO: 43, SEQ IDNO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53,SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO:58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ IDNO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ IDNO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42.

Methods for such manipulations are generally known in the art. Forexample, amino acid sequence variants of a PtIP-50 polypeptide orPtIP-65 polypeptide can be prepared by mutations in the DNA. This mayalso be accomplished by one of several forms of mutagenesis and/or indirected evolution. In some aspects, the changes encoded in the aminoacid sequence will not substantially affect the function of the protein.Such variants will possess the desired pesticidal activity. However, itis understood that the ability of a PtIP-50 polypeptide or PtIP-65polypeptide to confer pesticidal activity may be improved by the use ofsuch techniques upon the compositions of this disclosure.

For example, conservative amino acid substitutions may be made at one ormore, PtIP-, nonessential amino acid residues. A “nonessential” aminoacid residue is a residue that can be altered from the wild-typesequence of a PtIP-50 polypeptide or PtIP-65 polypeptide withoutaltering the biological activity. A “conservative amino acidsubstitution” is one in which the amino acid residue is replaced with anamino acid residue having a similar side chain. Families of amino acidresidues having similar side chains have been defined in the art. Thesefamilies include: amino acids with basic side chains (e.g., lysine,arginine, histidine); acidic side chains (e.g., aspartic acid, glutamicacid); polar, negatively charged residues and their amides (e.g.,aspartic acid, asparagine, glutamic, acid, glutamine; uncharged polarside chains (e.g., glycine, asparagine, glutamine, serine, threonine,tyrosine, cysteine); small aliphatic, nonpolar or slightly polarresidues (e.g., Alanine, serine, threonine, proline, glycine); nonpolarside chains (e.g., alanine, valine, leucine, isoleucine, proline,phenylalanine, methionine, tryptophan); large aliphatic, nonpolarresidues (e.g., methionine, leucine, isoleucine, valine, cystine);beta-branched side chains (e.g., threonine, valine, isoleucine);aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine); large aromatic side chains (e.g., tyrosine, phenylalanine,tryptophan).

Amino acid substitutions may be made in nonconserved regions that retainfunction. In general, such substitutions would not be made for conservedamino acid residues or for amino acid residues residing within aconserved motif, where such residues are essential for protein activity.Examples of residues that are conserved and that may be essential forprotein activity include, for example, residues that are identicalbetween all proteins contained in an alignment of similar or relatedtoxins to the sequences of the embodiments (e.g., residues that areidentical in an alignment of homologous proteins). Examples of residuesthat are conserved but that may allow conservative amino acidsubstitutions and still retain activity include, for example, residuesthat have only conservative substitutions between all proteins containedin an alignment of similar or related toxins to the sequences of theembodiments (e.g., residues that have only conservative substitutionsbetween all proteins contained in the alignment homologous proteins).However, one of skill in the art would understand that functionalvariants may have minor conserved or nonconserved alterations in theconserved residues. Guidance as to appropriate amino acid substitutionsthat do not affect biological activity of the protein of interest may befound in the model of Dayhoff, et al., (1978) Atlas of Protein Sequenceand Structure (Natl. Biomed. Res. Found., Washington, D.C.), hereinincorporated by reference.

In making such changes, the hydropathic index of amino acids may beconsidered. The importance of the hydropathic amino acid index inconferring interactive biologic function on a protein is generallyunderstood in the art (Kyte and Doolittle, (1982) J Mol Biol.157(1):105-32). It is accepted that the relative hydropathic characterof the amino acid contributes to the secondary structure of theresultant protein, which in turn defines the interaction of the proteinwith other molecules, for example, enzymes, substrates, receptors, DNA,antibodies, antigens, and the like.

It is known in the art that certain amino acids may be substituted byother amino acids having a similar hydropathic index or score and stillresult in a protein with similar biological activity, i.e., still obtaina biological functionally equivalent protein. Each amino acid has beenassigned a hydropathic index on the basis of its hydrophobicity andcharge characteristics (Kyte and Doolittle, ibid). These are: isoleucine(+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8);cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine(−0.4); threonine (−0.7); serine (−0.8); tryptophan (−0.9); tyrosine(−1.3); proline (−1.6); histidine (−3.2); glutamate (−3.5); glutamine(−3.5); aspartate (−3.5); asparagine (−3.5); lysine (−3.9) and arginine(−4.5). In making such changes, the substitution of amino acids whosehydropathic indices are within +2 is preferred, those which are within+1 are particularly preferred, and those within +0.5 are even moreparticularly preferred.

It is also understood in the art that the substitution of like aminoacids can be made effectively on the basis of hydrophilicity. U.S. Pat.No. 4,554,101, states that the greatest local average hydrophilicity ofa protein, as governed by the hydrophilicity of its adjacent aminoacids, correlates with a biological property of the protein.

As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicityvalues have been assigned to amino acid residues: arginine (+3.0);lysine (+3.0); aspartate (+3.0.+0.1); glutamate (+3.0.+0.1); serine(+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine(−0.4); proline (−0.5.+0.1); alanine (−0.5); histidine (−0.5); cysteine(−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine(−1.8); tyrosine (−2.3); phenylalanine (−2.5); tryptophan (−3.4).

Alternatively, alterations may be made to the protein sequence of manyproteins at the amino or carboxy terminus without substantiallyaffecting activity. This can include insertions, deletions oralterations introduced by modern molecular methods, such as PCR,including PCR amplifications that alter or extend the protein codingsequence by virtue of inclusion of amino acid encoding sequences in theoligonucleotides utilized in the PCR amplification. Alternatively, theprotein sequences added can include entire protein-coding sequences,such as those used commonly in the art to generate protein fusions. Suchfusion proteins are often used to (1) increase expression of a proteinof interest (2) introduce a binding domain, enzymatic activity orepitope to facilitate either protein purification, protein detection orother experimental uses known in the art (3) target secretion ortranslation of a protein to a subcellular organelle, such as theperiplasmic space of Gram-negative bacteria, mitochondria orchloroplasts of plants or the endoplasmic reticulum of eukaryotic cells,the latter of which often results in glycosylation of the protein.

Variant nucleotide and amino acid sequences of the disclosure alsoencompass sequences derived from mutagenic and recombinogenic proceduressuch as DNA shuffling. With such a procedure, one or more differentPtIP-50 polypeptide coding regions or one or more different PtIP-65polypeptide coding regions can be used to create a new PtIP-50polypeptide or PtIP-65 polypeptide possessing the desired properties. Inthis manner, libraries of recombinant polynucleotides are generated froma population of related sequence polynucleotides comprising sequenceregions that have substantial sequence identity and can be homologouslyrecombined in vitro or in vivo. For example, using this approach,sequence motifs encoding a domain of interest may be shuffled between apesticidal gene and other known pesticidal genes to obtain a new genecoding for a protein with an improved property of interest, such as anincreased insecticidal activity. Strategies for such DNA shuffling areknown in the art. See, for example, Stemmer, (1994) Proc. Natl. Acad.Sci. USA 91:10747-10751; Stemmer, (1994) Nature 370:389-391; Crameri, etal., (1997) Nature Biotech. 15:436-438; Moore, et al., (1997) J. Mol.Biol. 272:336-347; Zhang, et al., (1997) Proc. Natl. Acad. Sci. USA94:4504-4509; Crameri, et al., (1998) Nature 391:288-291; and U.S. Pat.Nos. 5,605,793 and 5,837,458.

Domain swapping or shuffling is another mechanism for generating alteredPtIP-50 polypeptides or PtIP-65 polypeptides. Domains may be swappedbetween PtIP-50 polypeptides or between PtIP-65 polypeptides, resultingin hybrid or chimeric toxins with improved insecticidal activity ortarget spectrum. Methods for generating recombinant proteins and testingthem for pesticidal activity are well known in the art (see, forexample, Naimov, et al., (2001) Appl. Environ. Microbiol. 67:5328-5330;de Maagd, et al., (1996) Appl. Environ. Microbiol. 62:1537-1543; Ge, etal., (1991) J. Biol. Chem. 266:17954-17958; Schnepf, et al., (1990) J.Biol. Chem. 265:20923-20930; Rang, et al., 91999) Appl. Environ.Microbiol. 65:2918-2925).

Alignment of PtIP-50 homologs (FIGS. 10a-10o, 11a-11f, 12a-12j, 13a-13e,14a-14e, 15a -15 e) allowed for identification of residues that arehighly conserved among natural homologs in this family. Alignment ofPtIP-65 homologs (FIGS. 3a-3i, 4a-4b, 5a-5c, 6a-6c , 7, 8 a-8 b) allowedfor identification of residues that are highly conserved among naturalhomologs in this family.

Compositions

Compositions comprising a PtIP-50 polypeptide and a PtIP-65 polypeptideof the disclosure are also embraced. In some embodiments the compositioncomprises a PtIP-50 polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQ IDNO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54,SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO:59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ IDNO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQID NO: 69 or SEQ ID NO: 70 and a PtIP-65 polypeptide of SEQ ID NO: 22,SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO:27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ IDNO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 orSEQ ID NO: 42. In some embodiments the composition comprises aPtIP-50/PtIP-65 fusion protein.

Antibodies

Antibodies to a PtIP-50 polypeptide or a PtIP-65 polypeptide of theembodiments or to variants or fragments thereof are also encompassed.The antibodies of the disclosure include polyclonal and monoclonalantibodies as well as fragments thereof which retain their ability tobind to PtIP-50 polypeptide or a PtIP-65 polypeptide found in the insectgut. An antibody, monoclonal antibody or fragment thereof is said to becapable of binding a molecule if it is capable of specifically reactingwith the molecule to thereby bind the molecule to the antibody,monoclonal antibody or fragment thereof. The term “antibody” (Ab) or“monoclonal antibody” (Mab) is meant to include intact molecules as wellas fragments or binding regions or domains thereof (such as, forexample, Fab and F(ab).sub.2 fragments) which are capable of bindinghapten. Such fragments are typically produced by proteolytic cleavage,such as papain or pepsin. Alternatively, hapten-binding fragments can beproduced through the application of recombinant DNA technology orthrough synthetic chemistry. Methods for the preparation of theantibodies of the present disclosure are generally known in the art. Forexample, see, Antibodies, A Laboratory Manual, Ed Harlow and David Lane(eds.) Cold Spring Harbor Laboratory, N.Y. (1988), as well as thereferences cited therein. Standard reference works setting forth thegeneral principles of immunology include: Klein, J. Immunology: TheScience of Cell-Noncell Discrimination, John Wiley & Sons, N.Y. (1982);Dennett, et al., Monoclonal Antibodies, Hybridoma: A New Dimension inBiological Analyses, Plenum Press, N.Y. (1980) and Campbell, “MonoclonalAntibody Technology,” In Laboratory Techniques in Biochemistry andMolecular Biology, Vol. 13, Burdon, et al., (eds.), Elsevier, Amsterdam(1984). See also, U.S. Pat. Nos. 4,196,265; 4,609,893; 4,713,325;4,714,681; 4,716,111; 4,716,117 and 4,720,459. PtIP-50 polypeptide orPtIP-65 polypeptide antibodies or antigen-binding portions thereof canbe produced by a variety of techniques, including conventionalmonoclonal antibody methodology, for example the standard somatic cellhybridization technique of Kohler and Milstein, (1975) Nature 256:495.Other techniques for producing monoclonal antibody can also be employedsuch as viral or oncogenic transformation of B lymphocytes. An animalsystem for preparing hybridomas is a murine system. Immunizationprotocols and techniques for isolation of immunized splenocytes forfusion are known in the art. Fusion partners (e.g., murine myelomacells) and fusion procedures are also known. The antibody and monoclonalantibodies of the disclosure can be prepared by utilizing a PtIP-50polypeptide or PtIP-65 polypeptide as antigens.

A kit for detecting the presence of a PtIP-50 polypeptide or a PtIP-65polypeptide or detecting the presence of a nucleotide sequence encodinga PtIP-50 polypeptide or a PtIP-65 polypeptide, in a sample is provided.In one embodiment, the kit provides antibody-based reagents fordetecting the presence of a PtIP-50 polypeptide or a PtIP-65 polypeptidein a tissue sample. In another embodiment, the kit provides labelednucleic acid probes useful for detecting the presence of one or morepolynucleotides encoding PtIP-50 polypeptide or a PtIP-65 polypeptide.The kit is provided along with appropriate reagents and controls forcarrying out a detection method, as well as instructions for use of thekit.

Receptor Identification and Isolation

Receptors to the PtIP-50 polypeptide and/or a PtIP-65 polypeptide of theembodiments or to variants or fragments thereof, are also encompassed.Methods for identifying receptors are well known in the art (see,Hofmann, et. al., (1988) Eur. J. Biochem. 173:85-91; Gill, et al.,(1995) J. Biol. Chem. 27277-27282) can be employed to identify andisolate the receptor that recognizes the PtIP-50 polypeptide and/or thePtIP-65 polypeptide using the brush-border membrane vesicles fromsusceptible insects. In addition to the radioactive labeling methodlisted in the cited literatures, PtIP-50 polypeptide or a PtIP-65polypeptide can be labeled with fluorescent dye and other common labelssuch as biotin. Brush-border membrane vesicles (BBMV) of susceptibleinsects such as soybean looper and stink bugs can be prepared accordingto the protocols listed in the references and separated on SDS-PAGE geland blotted on suitable membrane. Labeled PtIP-50 polypeptide or aPtIP-65 polypeptides can be incubated with blotted membrane of BBMV andlabeled the PtIP-50 polypeptide or a PtIP-65 polypeptides can beidentified with the labeled reporters. Identification of protein band(s)that interact with the PtIP-50 polypeptide or a PtIP-65 polypeptide canbe detected by N-terminal amino acid gas phase sequencing or massspectrometry based protein identification method (Patterson, (1998)10.22, 1-24, Current Protocol in Molecular Biology published by JohnWiley & Son Inc). Once the protein is identified, the corresponding genecan be cloned from genomic DNA or cDNA library of the susceptibleinsects and binding affinity can be measured directly with the PtIP-50polypeptide or a PtIP-65 polypeptide. Receptor function for insecticidalactivity by the PtIP-50 polypeptide or a PtIP-65 polypeptide can beverified by accomplished by RNAi type of gene knock out method(Rajagopal, et al., (2002) J. Biol. Chem. 277:46849-46851).

Nucleotide Constructs, Expression Cassettes and Vectors

The use of the term “nucleotide constructs” herein is not intended tolimit the embodiments to nucleotide constructs comprising DNA. Those ofordinary skill in the art will recognize that nucleotide constructsparticularly polynucleotides and oligonucleotides composed ofribonucleotides and combinations of ribonucleotides anddeoxyribonucleotides may also be employed in the methods disclosedherein. The nucleotide constructs, nucleic acids, and nucleotidesequences of the embodiments additionally encompass all complementaryforms of such constructs, molecules, and sequences. Further, thenucleotide constructs, nucleotide molecules, and nucleotide sequences ofthe embodiments encompass all nucleotide constructs, molecules, andsequences which can be employed in the methods of the embodiments fortransforming plants including, but not limited to, those comprised ofdeoxyribonucleotides, ribonucleotides, and combinations thereof. Suchdeoxyribonucleotides and ribonucleotides include both naturallyoccurring molecules and synthetic analogues. The nucleotide constructs,nucleic acids, and nucleotide sequences of the embodiments alsoencompass all forms of nucleotide constructs including, but not limitedto, single-stranded forms, double-stranded forms, hairpins,stem-and-loop structures and the like.

A further embodiment relates to a transformed organism such as anorganism selected from plant and insect cells, bacteria, yeast,baculovirus, protozoa, nematodes and algae. The transformed organismcomprises a DNA molecule of the embodiments, an expression cassettecomprising the DNA molecule or a vector comprising the expressioncassette, which may be stably incorporated into the genome of thetransformed organism.

The sequences of the embodiments are provided in DNA constructs forexpression in the organism of interest. The construct will include 5′and 3′ regulatory sequences operably linked to a sequence of theembodiments. The term “operably linked” as used herein refers to afunctional linkage between a promoter and a second sequence, wherein thepromoter sequence initiates and mediates transcription of the DNAsequence corresponding to the second sequence. Generally, operablylinked means that the nucleic acid sequences being linked are contiguousand where necessary to join two protein coding regions in the samereading frame. The construct may additionally contain at least oneadditional gene to be cotransformed into the organism. Alternatively,the additional gene(s) can be provided on multiple DNA constructs.

Such a DNA construct is provided with a plurality of restriction sitesfor insertion of the PtIP-50 polypeptide or a PtIP-65 polypeptide genesequence to be under the transcriptional regulation of the regulatoryregions. The DNA construct may additionally contain selectable markergenes.

The DNA construct will generally include in the 5′ to 3′ direction oftranscription: a transcriptional and translational initiation region(i.e., a promoter), a DNA sequence of the embodiments, and atranscriptional and translational termination region (i.e., terminationregion) functional in the organism serving as a host. Thetranscriptional initiation region (i.e., the promoter) may be native,analogous, foreign or heterologous to the host organism and/or to thesequence of the embodiments. Additionally, the promoter may be thenatural sequence or alternatively a synthetic sequence. The term“foreign” as used herein indicates that the promoter is not found in thenative organism into which the promoter is introduced. Where thepromoter is “foreign” or “heterologous” to the sequence of theembodiments, it is intended that the promoter is not the native ornaturally occurring promoter for the operably linked sequence of theembodiments. As used herein, a chimeric gene comprises a coding sequenceoperably linked to a transcription initiation region that isheterologous to the coding sequence. Where the promoter is a native ornatural sequence, the expression of the operably linked sequence isaltered from the wild-type expression, which results in an alteration inphenotype.

In some embodiments the DNA construct may also include a transcriptionalenhancer sequence. As used herein, the term an “enhancer” refers to aDNA sequence which can stimulate promoter activity, and may be an innateelement of the promoter or a heterologous element inserted to enhancethe level or tissue-specificity of a promoter. Various enhancers areknown in the art including for example, introns with gene expressionenhancing properties in plants (US Patent Application Publication Number2009/0144863, the ubiquitin intron (i.e., the maize ubiquitin intron 1(see, for example, NCBI sequence S94464)), the omega enhancer or theomega prime enhancer (Gallie, et al., (1989) Molecular Biology of RNAed. Cech (Liss, New York) 237-256 and Gallie, et al., (1987) Gene60:217-25), the CaMV 35S enhancer (see, e.g., Benfey, et al., (1990)EMBO J. 9:1685-96) and the enhancers of U.S. Pat. No. 7,803,992 may alsobe used, each of which is incorporated by reference. The above list oftranscriptional enhancers is not meant to be limiting. Any appropriatetranscriptional enhancer can be used in the embodiments.

The termination region may be native with the transcriptional initiationregion, may be native with the operably linked DNA sequence of interest,may be native with the plant host or may be derived from another source(i.e., foreign or heterologous to the promoter, the sequence ofinterest, the plant host or any combination thereof). Convenienttermination regions are available from the Ti-plasmid of A. tumefaciens,such as the octopine synthase and nopaline synthase termination regions.See also, Guerineau, et al., (1991) Mol. Gen. Genet. 262:141-144;Proudfoot, (1991) Cell 64:671-674; Sanfacon, et al., (1991) Genes Dev.5:141-149; Mogen, et al., (1990) Plant Cell 2:1261-1272; Munroe, et al.,(1990) Gene 91:151-158; Ballas, et al., (1989) Nucleic Acids Res.17:7891-7903 and Joshi, et al., (1987) Nucleic Acid Res. 15:9627-9639.

Where appropriate, a nucleic acid may be optimized for increasedexpression in the host organism. Thus, where the host organism is aplant, the synthetic nucleic acids can be synthesized usingplant-preferred codons for improved expression. See, for example,Campbell and Gown, (1990) Plant Physiol. 92:1-11 for a discussion ofhost-preferred codon usage. For example, although nucleic acid sequencesof the embodiments may be expressed in both monocotyledonous anddicotyledonous plant species, sequences can be modified to account forthe specific codon preferences and GC content preferences ofmonocotyledons or dicotyledons as these preferences have been shown todiffer (Murray et al. (1989) Nucleic Acids Res. 17:477-498). Thus, themaize-preferred codon for a particular amino acid may be derived fromknown gene sequences from maize. Maize codon usage for 28 genes frommaize plants is listed in Table 4 of Murray, et al., supra. Methods areavailable in the art for synthesizing plant-preferred genes. See, forexample, U.S. Pat. Nos. 5,380,831, and 5,436,391 and Murray, et al.,(1989) Nucleic Acids Res. 17:477-498, and Liu H et al. Mol Bio Rep37:677-684, 2010, herein incorporated by reference. A Zea maize codonusage table can be also found atkazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=4577, which can beaccessed using the www prefix. Table 4 shows a maize optimal codonanalysis (adapted from Liu H et al. Mol Bio Rep 37:677-684, 2010).

TABLE 4 Amino High Low Amino High Low Acid Codon Count RSCU Count RSCUAcid Codon Count RSCU Count RSCU Phe UUU 115 0.04 2,301 1.22 Ala GCU 6290.17 3,063 1.59 UUC* 5,269 1.96 1,485 0.78 GCC* 8,057 2.16 1,136 0.59Ser UCU 176 0.13 2,498 1.48 GCA 369 0.1 2,872 1.49 UCC* 3,489 2.48 1,0740.63 GCG* 5,835 1.57 630 0.33 UCA 104 0.07 2,610 1.54 Tyr UAU 71 0.041,632 1.22 UCG* 1,975 1.4 670 0.4 UAC* 3,841 1.96 1,041 0.78 AGU 77 0.051,788 1.06 His CAU 131 0.09 1,902 1.36 AGC* 2,617 1.86 1,514 0.89 CAC*2,800 1.91 897 0.64 Leu UUA 10 0.01 1,326 0.79 Cys UGU 52 0.04 1,2331.12 UUG 174 0.09 2,306 1.37 UGC* 2,291 1.96 963 0.88 CUU 223 0.11 2,3961.43 Gln CAA 99 0.05 2,312 1.04 CUC* 5,979 3.08 1,109 0.66 CAG* 3,5571.95 2,130 0.96 CUA 106 0.05 1,280 0.76 Arg CGU 153 0.12 751 0.74 CUG*5,161 2.66 1,646 0.98 CGC* 4,278 3.25 466 0.46 Pro CCU 427 0.22 1,9001.47 CGA 92 0.07 659 0.65 CCC* 3,035 1.59 601 0.47 CGG* 1,793 1.36 6310.62 CCA 311 0.16 2,140 1.66 AGA 83 0.06 1,948 1.91 CCG* 3,846 2.02 5130.4 AGG* 1,493 1.14 1,652 1.62 Ile AUU 138 0.09 2,388 1.3 Asn AAU 1310.07 3,074 1.26 AUC* 4,380 2.85 1,353 0.74 AAC* 3,814 1.93 1,807 0.74AUA 88 0.06 1,756 0.96 Lys AAA 130 0.05 3,215 0.98 Thr ACU 136 0.091,990 1.43 AAG* 5,047 1.95 3,340 1.02 ACC* 3,398 2.25 991 0.71 Asp GAU312 0.09 4,217 1.38 ACA 133 0.09 2,075 1.5 GAC* 6,729 1.91 1,891 0.62ACG* 2,378 1.57 495 0.36 Gly GGU 363 0.13 2,301 1.35 Val GUU 182 0.072,595 1.51 GGC* 7,842 2.91 1,282 0.75 GUC* 4,584 1.82 1,096 0.64 GGA 3970.15 2,044 1.19 GUA 74 0.03 1,325 0.77 GGG* 2,186 0.81 1,215 0.71 GUG*5,257 2.08 1,842 1.07 Glu GAA 193 0.06 4,080 1.1 GAG* 6,010 1.94 3,3070.9 Codon usage was compared using Chi squared contingency test toidentify optimal codons. Codons that occur significantly more often(P\0.01) are indicated with an asterisk.

A Glycine max codon usage table is shown in Table 5 and can also befound atkazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=3847&aa=1&style=N,which can be accessed using the www prefix.

TABLE 5 TTT F 21.2 (10493) TCT S 18.4 (9107) TTC F 21.2 (10487) TCC S12.9 (6409) TTA L 9.2  (4545) TCA S 15.6 (7712) TTG L 22.9 (11340) TCG S4.8 (2397) CTT L 23.9 (11829) CCT P 18.9 (9358) CTC L 17.1  (8479) CCC P10.1 (5010) CTA L 8.5  (4216) CCA P 19.1 (9461) CTG L 12.7  (6304) CCG P4.7 (2312) ATT I 25.1 (12411) ACT T 17.1 (8490) ATC I 16.3  (8071) ACC T14.3 (7100) ATA I 12.9  (6386) ACA T 14.9 (7391) ATG M 22.7 (11218) ACGT 4.3 (2147) GTT V 26.1 (12911) GCT A 26.7 (13201)  GTC V 11.9  (5894)GCC A 16.2 (8026) GTA V 7.7  (3803) GCA A 21.4 (10577)  GTG V 21.4(10610) GCG A 6.3 (3123) TAT Y 15.7  (7779) TGT C 8.1 (3995) TAC Y 14.9 (7367) TGC C 8.0 (3980) TAA * 0.9  (463) TGA * 1.0  (480) TAG * 0.5 (263) TGG W 13.0 (6412) CAT H 14.0  (6930) CGT R 6.6 (3291) CAC H 11.6 (5759) CGC R 6.2 (3093) CAA Q 20.5 (10162) CGA R 4.1 (2018) CAG Q 16.2 (8038) CGG R 3.1 (1510) AAT N 22.4 (11088) AGT S 12.6 (6237) AAC N 22.8(11284) AGC S 11.3 (5594) AAA K 26.9 (13334) AGA R 14.8 (7337) AAG K35.9 (17797) AGG R 13.3 (6574) GAT D 32.4 (16040) GGT G 20.9 (10353) GAC D 20.4 (10097) GGC G 13.4 (6650) GAA E 33.2 (16438) GGA G 22.3(11022)  GAG E 33.2 (16426) GGG G 13.0 (6431)

In some embodiments the recombinant nucleic acid molecule encoding aPtIP-50 polypeptide or a PtIP-65 polypeptide has maize optimized codons.

Additional sequence modifications are known to enhance gene expressionin a cellular host. These include elimination of sequences encodingspurious polyadenylation signals, exon-intron splice site signals,transposon-like repeats, and other well-characterized sequences that maybe deleterious to gene expression. The GC content of the sequence may beadjusted to levels average for a given cellular host, as calculated byreference to known genes expressed in the host cell. The term “hostcell” as used herein refers to a cell which contains a vector andsupports the replication and/or expression of the expression vector isintended. Host cells may be prokaryotic cells such as E. coli oreukaryotic cells such as yeast, insect, amphibian or mammalian cells ormonocotyledonous or dicotyledonous plant cells. An example of amonocotyledonous host cell is a maize host cell. When possible, thesequence is modified to avoid PtIP-hairpin secondary mRNA structures.

The expression cassettes may additionally contain 5′ leader sequences.Such leader sequences can act to enhance translation. Translationleaders are known in the art and include: picornavirus leaders, forexample, EMCV leader (Encephalomyocarditis 5′ noncoding region)(Elroy-Stein, et al., (1989) Proc. Natl. Acad. Sci. USA 86:6126-6130);potyvirus leaders, for example, TEV leader (Tobacco Etch Virus) (Gallie,et al., (1995) Gene 165(2):233-238), MDMV leader (Maize Dwarf MosaicVirus), human immunoglobulin heavy-chain binding protein (BiP) (Macejak,et al., (1991) Nature 353:90-94); untranslated leader from the coatprotein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling, et al.,(1987) Nature 325:622-625); tobacco mosaic virus leader (TMV) (Gallie,et al., (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York),pp. 237-256) and maize chlorotic mottle virus leader (MCMV) (Lommel, etal., (1991) Virology 81:382-385). See also, Della-Cioppa, et al., (1987)Plant Physiol. 84:965-968. Such constructs may also contain a “signalsequence” or “leader sequence” to facilitate co-translational orpost-translational transport of the peptide to certain intracellularstructures such as the chloroplast (or other plastid), endoplasmicreticulum or Golgi apparatus.

“Signal sequence” as used herein refers to a sequence that is known orsuspected to result in cotranslational or post-translational peptidetransport across the cell membrane. In eukaryotes, this typicallyinvolves secretion into the Golgi apparatus, with some resultingglycosylation. Insecticidal toxins of bacteria are often synthesized asprotoxins, which are protolytically activated in the gut of the targetpest (Chang, (1987) Methods Enzymol. 153:507-516). In some embodiments,the signal sequence is located in the native sequence or may be derivedfrom a sequence of the embodiments. “Leader sequence” as used hereinrefers to any sequence that when translated, results in an amino acidsequence sufficient to trigger co-translational transport of the peptidechain to a subcellular organelle. Thus, this includes leader sequencestargeting transport and/or glycosylation by passage into the endoplasmicreticulum, passage to vacuoles, plastids including chloroplasts,mitochondria, and the like. Nuclear-encoded proteins targeted to thechloroplast thylakoid lumen compartment have a characteristic bipartitetransit peptide, composed of a stromal targeting signal peptide and alumen targeting signal peptide. The stromal targeting information is inthe amino-proximal portion of the transit peptide. The lumen targetingsignal peptide is in the carboxyl-proximal portion of the transitpeptide, and contains all the information for targeting to the lumen.Recent research in proteomics of the higher plant chloroplast hasachieved in the identification of numerous nuclear-encoded lumenproteins (Kieselbach et al. FEBS LETT 480:271-276, 2000; Peltier et al.Plant Cell 12:319-341, 2000; Bricker et al. Biochim. Biophys Acta1503:350-356, 2001), the lumen targeting signal peptide of which canpotentially be used in accordance with the present disclosure. About 80proteins from Arabidopsis, as well as homologous proteins from spinachand garden pea, are reported by Kieselbach et al., PhotosynthesisResearch, 78:249-264, 2003. In particular, Table 2 of this publication,which is incorporated into the description herewith by reference,discloses 85 proteins from the chloroplast lumen, identified by theiraccession number (see also US Patent Application Publication2009/09044298). In addition, the recently published draft version of therice genome (Goff et al, Science 296:92-100, 2002) is a suitable sourcefor lumen targeting signal peptide which may be used in accordance withthe present disclosure.

Suitable chloroplast transit peptides (CTP) are well known to oneskilled in the art also include chimeric CTPs comprising but not limitedto, an N-terminal domain, a central domain or a C-terminal domain from aCTP from Oryza sativa 1-deoxy-D xyulose-5-Phosphate Synthase Oryzasativa-Superoxide dismutase Oryza sativa-soluble starch synthase Oryzasativa-NADP-dependent Malic acid enzyme Oryzasativa-Phospho-2-dehydro-3-deoxyheptonate Aldolase 2 Oryzasativa-L-Ascorbate peroxidase 5 Oryza sativa-Phosphoglucan waterdikinase, Zea Mays ssRUBISCO, Zea Mays-beta-glucosidase, Zea Mays-Malatedehydrogenase, Zea Mays Thioredoxin M-type US Patent ApplicationPublication 2012/0304336).

The PtIP-50 polypeptide or a PtIP-65 polypeptide gene to be targeted tothe chloroplast may be optimized for expression in the chloroplast toaccount for differences in codon usage between the plant nucleus andthis organelle. In this manner, the nucleic acids of interest may besynthesized using chloroplast-preferred codons. See, for example, U.S.Pat. No. 5,380,831, herein incorporated by reference.

In preparing the expression cassette, the various DNA fragments may bemanipulated so as to provide for the DNA sequences in the properorientation and, as appropriate, in the proper reading frame. Towardthis end, adapters or linkers may be employed to join the DNA fragmentsor other manipulations may be involved to provide for convenientrestriction sites, removal of superfluous DNA, removal of restrictionsites or the like. For this purpose, in vitro mutagenesis, primerrepair, restriction, annealing, resubstitutions, e.g., transitions andtransversions, may be involved.

A number of promoters can be used in the practice of the embodiments.The promoters can be selected based on the desired outcome. The nucleicacids can be combined with constitutive, tissue-preferred, inducible orother promoters for expression in the host organism. Suitableconstitutive promoters for use in a plant host cell include, forexample, the core promoter of the Rsyn7 promoter and other constitutivepromoters disclosed in WO 1999/43838 and U.S. Pat. No. 6,072,050; thecore CaMV 35S promoter (Odell, et al., (1985) Nature 313:810-812); riceactin (McElroy, et al., (1990) Plant Cell 2:163-171); ubiquitin(Christensen, et al., (1989) Plant Mol. Biol. 12:619-632 andChristensen, et al., (1992) Plant Mol. Biol. 18:675-689); pEMU (Last, etal., (1991) Theor. Appl. Genet. 81:581-588); MAS (Velten, et al., (1984)EMBO J. 3:2723-2730); ALS promoter (U.S. Pat. No. 5,659,026) and thelike. Other constitutive promoters include, for example, those discussedin U.S. Pat. Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785;5,399,680; 5,268,463; 5,608,142 and 6,177,611.

Depending on the desired outcome, it may be beneficial to express thegene from an inducible promoter. Of particular interest for regulatingthe expression of the nucleotide sequences of the embodiments in plantsare wound-inducible promoters. Such wound-inducible promoters, mayrespond to damage caused by insect feeding, and include potatoproteinase inhibitor (pin II) gene (Ryan, (1990) Ann. Rev. Phytopath.28:425-449; Duan, et al., (1996) Nature Biotechnology 14:494-498); wun1and wun2, U.S. Pat. No. 5,428,148; win1 and win2 (Stanford, et al.,(1989) Mol. Gen. Genet. 215:200-208); systemin (McGurl, et al., (1992)Science 225:1570-1573); WIP1 (Rohmeier, et al., (1993) Plant Mol. Biol.22:783-792; Eckelkamp, et al., (1993) FEBS Letters 323:73-76); MPI gene(Corderok, et al., (1994) Plant J. 6(2):141-150) and the like, hereinincorporated by reference.

Additionally, pathogen-inducible promoters may be employed in themethods and nucleotide constructs of the embodiments. Suchpathogen-inducible promoters include those from pathogenesis-relatedproteins (PR proteins), which are induced following infection by apathogen; e.g., PR proteins, SAR proteins, beta-1,3-glucanase,chitinase, etc. See, for example, Redolfi, et al., (1983) Neth. J. PlantPathol. 89:245-254; Uknes, et al., (1992) Plant Cell 4: 645-656 and VanLoon, (1985) Plant Mol. Virol. 4:111-116. See also, WO 1999/43819,herein incorporated by reference.

Of interest are promoters that are expressed locally at or near the siteof pathogen infection. See, for example, Marineau, et al., (1987) PlantMol. Biol. 9:335-342; Matton, et al., (1989) Molecular Plant-MicrobeInteractions 2:325-331; Somsisch, et al., (1986) Proc. Natl. Acad. Sci.USA 83:2427-2430; Somsisch, et al., (1988) Mol. Gen. Genet. 2:93-98 andYang, (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977. See also, Chen,et al., (1996) Plant J. 10:955-966; Zhang, et al., (1994) Proc. Natl.Acad. Sci. USA 91:2507-2511; Warner, et al., (1993) Plant J. 3:191-201;Siebertz, et al., (1989) Plant Cell 1:961-968; U.S. Pat. No. 5,750,386(nematode-inducible) and the references cited therein. Of particularinterest is the inducible promoter for the maize PRms gene, whoseexpression is induced by the pathogen Fusarium moniliforme (see, forexample, Cordero, et al., (1992) Physiol. Mol. Plant Path. 41:189-200).

Chemical-regulated promoters can be used to modulate the expression of agene in a plant through the application of an exogenous chemicalregulator. Depending upon the objective, the promoter may be achemical-inducible promoter, where application of the chemical inducesgene expression or a chemical-repressible promoter, where application ofthe chemical represses gene expression. Chemical-inducible promoters areknown in the art and include, but are not limited to, the maize In2-2promoter, which is activated by benzenesulfonamide herbicide safeners,the maize GST promoter, which is activated by hydrophobic electrophiliccompounds that are used as pre-emergent herbicides, and the tobaccoPR-la promoter, which is activated by salicylic acid. Otherchemical-regulated promoters of interest include steroid-responsivepromoters (see, for example, the glucocorticoid-inducible promoter inSchena, et al., (1991) Proc. Natl. Acad. Sci. USA 88:10421-10425 andMcNellis, et al., (1998) Plant J. 14(2):247-257) andtetracycline-inducible and tetracycline-repressible promoters (see, forexample, Gatz, et al., (1991) Mol. Gen. Genet. 227:229-237 and U.S. Pat.Nos. 5,814,618 and 5,789,156), herein incorporated by reference.

Tissue-preferred promoters can be utilized to target enhanced PtIP-50polypeptide or a PtIP-65 polypeptide expression within a particularplant tissue. Tissue-preferred promoters include those discussed inYamamoto, et al., (1997) Plant J. 12(2)255-265; Kawamata, et al., (1997)Plant Cell Physiol. 38(7):792-803; Hansen, et al., (1997) Mol. GenGenet. 254(3):337-343; Russell, et al., (1997) Transgenic Res.6(2):157-168; Rinehart, et al., (1996) Plant Physiol. 112(3):1331-1341;Van Camp, et al., (1996) Plant Physiol. 112(2):525-535; Canevascini, etal., (1996) Plant Physiol. 112(2):513-524; Yamamoto, et al., (1994)Plant Cell Physiol. 35(5):773-778; Lam, (1994) Results Probl. CellDiffer. 20:181-196; Orozco, et al., (1993) Plant Mol Biol.23(6):1129-1138; Matsuoka, et al., (1993) Proc Natl. Acad. Sci. USA90(20):9586-9590 and Guevara-Garcia, et al., (1993) Plant J.4(3):495-505. Such promoters can be modified, if necessary, for weakexpression.

Leaf-preferred promoters are known in the art. See, for example,Yamamoto, et al., (1997) Plant J. 12(2):255-265; Kwon, et al., (1994)Plant Physiol. 105:357-67; Yamamoto, et al., (1994) Plant Cell Physiol.35(5):773-778; Gotor, et al., (1993) Plant J. 3:509-18; Orozco, et al.,(1993) Plant Mol. Biol. 23(6):1129-1138 and Matsuoka, et al., (1993)Proc. Natl. Acad. Sci. USA 90(20):9586-9590.

Root-preferred or root-specific promoters are known and can be selectedfrom the many available from the literature or isolated de novo fromvarious compatible species. See, for example, Hire, et al., (1992) PlantMol. Biol. 20(2):207-218 (soybean root-specific glutamine synthetasegene); Keller and Baumgartner, (1991) Plant Cell 3(10):1051-1061(root-specific control element in the GRP 1.8 gene of French bean);Sanger, et al., (1990) Plant Mol. Biol. 14(3):433-443 (root-specificpromoter of the mannopine synthase (MAS) gene of Agrobacteriumtumefaciens) and Miao, et al., (1991) Plant Cell 3(1):11-22 (full-lengthcDNA clone encoding cytosolic glutamine synthetase (GS), which isexpressed in roots and root nodules of soybean). See also, Bogusz, etal., (1990) Plant Cell 2(7):633-641, where two root-specific promotersisolated from hemoglobin genes from the nitrogen-fixing nonlegumeParasponia andersonii and the related non-nitrogen-fixing nonlegumeTrema tomentosa are described. The promoters of these genes were linkedto a β-glucuronidase reporter gene and introduced into both thenonlegume Nicotiana tabacum and the legume Lotus corniculatus, and inboth instances root-specific promoter activity was preserved. Leach andAoyagi, (1991) describe their analysis of the promoters of the highlyexpressed rolC and rolD root-inducing genes of Agrobacterium rhizogenes(see, Plant Science (Limerick) 79(1):69-76). They concluded thatenhancer and tissue-preferred DNA determinants are dissociated in thosepromoters. Teeri, et al., (1989) used gene fusion to lacZ to show thatthe Agrobacterium T-DNA gene encoding octopine synthase is especiallyactive in the epidermis of the root tip and that the TR2′ gene is rootspecific in the intact plant and stimulated by wounding in leaf tissue,an especially desirable combination of characteristics for use with aninsecticidal or larvicidal gene (see, EMBO J. 8(2):343-350). The TR1′gene fused to nptII (neomycin phosphotransferase II) showed similarcharacteristics. Additional root-preferred promoters include theVfENOD-GRP3 gene promoter (Kuster, et al., (1995) Plant Mol. Biol.29(4):759-772) and rolB promoter (Capana, et al., (1994) Plant Mol.Biol. 25(4):681-691). See also, U.S. Pat. Nos. 5,837,876; 5,750,386;5,633,363; 5,459,252; 5,401,836; 5,110,732 and 5,023,179. Arabidopsisthaliana root-preferred regulatory sequences are disclosed inUS20130117883.

“Seed-preferred” promoters include both “seed-specific” promoters (thosepromoters active during seed development such as promoters of seedstorage proteins) as well as “seed-germinating” promoters (thosepromoters active during seed germination). See, Thompson, et al., (1989)BioEssays 10:108, herein incorporated by reference. Such seed-preferredpromoters include, but are not limited to, Cim1 (cytokinin-inducedmessage); cZ19B1 (maize 19 kDa zein); and milps(myo-inositol-1-phosphate synthase) (see, U.S. Pat. No. 6,225,529,herein incorporated by reference). Gamma-zein and Glb-1 areendosperm-specific promoters. For dicots, seed-specific promotersinclude, but are not limited to, Kunitz trypsin inhibitor 3 (KTi3)(Jofuku and Goldberg, (1989) Plant Cell 1:1079-1093), bean β-phaseolin,napin, β-conglycinin, glycinin 1, soybean lectin, cruciferin, and thelike. For monocots, seed-specific promoters include, but are not limitedto, maize 15 kDa zein, 22 kDa zein, 27 kDa zein, g-zein, waxy, shrunken1, shrunken 2, globulin 1, etc. See also, WO 2000/12733, whereseed-preferred promoters from end1 and end2 genes are disclosed; hereinincorporated by reference. In dicots, seed specific promoters includebut are not limited to seed coat promoter from Arabidopsis, pBAN; andthe early seed promoters from Arabidopsis, p26, p63, and p63tr (U.S.Pat. Nos. 7,294,760 and 7,847,153). A promoter that has “preferred”expression in a particular tissue is expressed in that tissue to agreater degree than in at least one other plant tissue. Sometissue-preferred promoters show expression almost exclusively in theparticular tissue.

Where low level expression is desired, weak promoters will be used.Generally, the term “weak promoter” as used herein refers to a promoterthat drives expression of a coding sequence at a low level. By low levelexpression at levels of between about 1/1000 transcripts to about1/100,000 transcripts to about 1/500,000 transcripts is intended.Alternatively, it is recognized that the term “weak promoters” alsoencompasses promoters that drive expression in only a few cells and notin others to give a total low level of expression. Where a promoterdrives expression at unacceptably high levels, portions of the promotersequence can be deleted or modified to decrease expression levels.

Such weak constitutive promoters include, for example the core promoterof the Rsyn7 promoter (WO 1999/43838 and U.S. Pat. No. 6,072,050), thecore 35S CaMV promoter, and the like. Other constitutive promotersinclude, for example, those disclosed in U.S. Pat. Nos. 5,608,149;5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463;5,608,142 and 6,177,611, herein incorporated by reference.

The above list of promoters is not meant to be limiting. Any appropriatepromoter can be used in the embodiments.

Generally, the expression cassette will comprise a selectable markergene for the selection of transformed cells. Selectable marker genes areutilized for the selection of transformed cells or tissues. Marker genesinclude genes encoding antibiotic resistance, such as those encodingneomycin phosphotransferase II (NEO) and hygromycin phosphotransferase(HPT), as well as genes conferring resistance to herbicidal compounds,such as glufosinate ammonium, bromoxynil, imidazolinones and2,4-dichlorophenoxyacetate (2,4-D). Additional examples of suitableselectable marker genes include, but are not limited to, genes encodingresistance to chloramphenicol (Herrera Estrella, et al., (1983) EMBO J.2:987-992); methotrexate (Herrera Estrella, et al., (1983) Nature303:209-213 and Meijer, et al., (1991) Plant Mol. Biol. 16:807-820);streptomycin (Jones, et al., (1987) Mol. Gen. Genet. 210:86-91);spectinomycin (Bretagne-Sagnard, et al., (1996) Transgenic Res.5:131-137); bleomycin (Hille, et al., (1990) Plant Mol. Biol.7:171-176); sulfonamide (Guerineau, et al., (1990) Plant Mol. Biol.15:127-136); bromoxynil (Stalker, et al., (1988) Science 242:419-423);glyphosate (Shaw, et al., (1986) Science 233:478-481 and U.S. patentapplication Ser. Nos. 10/004,357 and 10/427,692); phosphinothricin(DeBlock, et al., (1987) EMBO J. 6:2513-2518). See generally, Yarranton,(1992) Curr. Opin. Biotech. 3:506-511; Christopherson, et al., (1992)Proc. Natl. Acad. Sci. USA 89:6314-6318; Yao, et al., (1992) Cell71:63-72; Reznikoff, (1992) Mol. Microbiol. 6:2419-2422; Barkley, etal., (1980) in The Operon, pp. 177-220; Hu, et al., (1987) Cell48:555-566; Brown, et al., (1987) Cell 49:603-612; Figge, et al., (1988)Cell 52:713-722; Deuschle, et al., (1989) Proc. Natl. Acad. Sci. USA86:5400-5404; Fuerst, et al., (1989) Proc. Natl. Acad. Sci. USA86:2549-2553; Deuschle, et al., (1990) Science 248:480-483; Gossen,(1993) Ph.D. Thesis, University of Heidelberg; Reines, et al., (1993)Proc. Natl. Acad. Sci. USA 90:1917-1921; Labow, et al., (1990) Mol.Cell. Biol. 10:3343-3356; Zambretti, et al., (1992) Proc. Natl. Acad.Sci. USA 89:3952-3956; Baim, et al., (1991) Proc. Natl. Acad. Sci. USA88:5072-5076; Wyborski, et al., (1991) Nucleic Acids Res. 19:4647-4653;Hillenand-Wissman, (1989) Topics Mol. Struc. Biol. 10:143-162;Degenkolb, et al., (1991) Antimicrob. Agents Chemother. 35:1591-1595;Kleinschnidt, et al., (1988) Biochemistry 27:1094-1104; Bonin, (1993)Ph.D. Thesis, University of Heidelberg; Gossen, et al., (1992) Proc.Natl. Acad. Sci. USA 89:5547-5551; Oliva, et al., (1992) Antimicrob.Agents Chemother. 36:913-919; Hlavka, et al., (1985) Handbook ofExperimental Pharmacology, Vol. 78 (Springer-Verlag, Berlin) and Gill,et al., (1988) Nature 334:721-724. Such disclosures are hereinincorporated by reference.

The above list of selectable marker genes is not meant to be limiting.Any selectable marker gene can be used in the embodiments.

Plant Transformation

The methods of the embodiments involve introducing a polypeptide orpolynucleotide into a plant. “Introducing” is as used herein meanspresenting to the plant the polynucleotide or polypeptide in such amanner that the sequence gains access to the interior of a cell of theplant. The methods of the embodiments do not depend on a particularmethod for introducing a polynucleotide or polypeptide into a plant,only that the polynucleotide or polypeptides gains access to theinterior of at least one cell of the plant. Methods for introducingpolynucleotide or polypeptides into plants are known in the artincluding, but not limited to, stable transformation methods, transienttransformation methods, and virus-mediated methods.

“Stable transformation” is as used herein means that the nucleotideconstruct introduced into a plant integrates into the genome of theplant and is capable of being inherited by the progeny thereof.“Transient transformation” as used herein means that a polynucleotide isintroduced into the plant and does not integrate into the genome of theplant or a polypeptide is introduced into a plant. “Plant” as usedherein refers to whole plants, plant organs (e.g., leaves, stems, roots,etc.), seeds, plant cells, propagules, embryos and progeny of the same.Plant cells can be differentiated or undifferentiated (e.g. callus,suspension culture cells, protoplasts, leaf cells, root cells, phloemcells and pollen).

Transformation protocols as well as protocols for introducing nucleotidesequences into plants may vary depending on the type of plant or plantcell, i.e., monocot or dicot, targeted for transformation. Suitablemethods of introducing nucleotide sequences into plant cells andsubsequent insertion into the plant genome include microinjection(Crossway, et al., (1986) Biotechniques 4:320-334), electroporation(Riggs, et al., (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606),Agrobacterium-mediated transformation (U.S. Pat. Nos. 5,563,055 and5,981,840), direct gene transfer (Paszkowski, et al., (1984) EMBO J.3:2717-2722) and ballistic particle acceleration (see, for example, U.S.Pat. Nos. 4,945,050; 5,879,918; 5,886,244 and 5,932,782; Tomes, et al.,(1995) in Plant Cell, Tissue, and Organ Culture: Fundamental Methods,ed. Gamborg and Phillips, (Springer-Verlag, Berlin) and McCabe, et al.,(1988) Biotechnology 6:923-926) and Lecl transformation (WO 00/28058).For potato transformation see, Tu, et al., (1998) Plant MolecularBiology 37:829-838 and Chong, et al., (2000) Transgenic Research9:71-78. Additional transformation procedures can be found inWeissinger, et al., (1988) Ann. Rev. Genet. 22:421-477; Sanford, et al.,(1987) Particulate Science and Technology 5:27-37 (onion); Christou, etal., (1988) Plant Physiol. 87:671-674 (soybean); McCabe, et al., (1988)Bio/Technology 6:923-926 (soybean); Finer and McMullen, (1991) In VitroCell Dev. Biol. 27P:175-182 (soybean); Singh, et al., (1998) Theor.Appl. Genet. 96:319-324 (soybean); Datta, et al., (1990) Biotechnology8:736-740 (rice); Klein, et al., (1988) Proc. Natl. Acad. Sci. USA85:4305-4309 (maize); Klein, et al., (1988) Biotechnology 6:559-563(maize); U.S. Pat. Nos. 5,240,855; 5,322,783 and 5,324,646; Klein, etal., (1988) Plant Physiol. 91:440-444 (maize); Fromm, et al., (1990)Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren, et al., (1984)Nature (London) 311:763-764; U.S. Pat. No. 5,736,369 (cereals);Bytebier, et al., (1987) Proc. Natl. Acad. Sci. USA 84:5345-5349(Liliaceae); De Wet, et al., (1985) in The Experimental Manipulation ofOvule Tissues, ed. Chapman, et al., (Longman, New York), pp. 197-209(pollen); Kaeppler, et al., (1990) Plant Cell Reports 9:415-418 andKaeppler, et al., (1992) Theor. Appl. Genet. 84:560-566(whisker-mediated transformation); D'Halluin, et al., (1992) Plant Cell4:1495-1505 (electroporation); Li, et al., (1993) Plant Cell Reports12:250-255 and Christou and Ford, (1995) Annals of Botany 75:407-413(rice); Osjoda, et al., (1996) Nature Biotechnology 14:745-750 (maizevia Agrobacterium tumefaciens); all of which are herein incorporated byreference.

In specific embodiments, the sequences of the embodiments can beprovided to a plant using a variety of transient transformation methods.Such transient transformation methods include, but are not limited to,the introduction of the PtIP-50 polypeptide or a PtIP-65 polypeptide orvariants and fragments thereof directly into the plant or theintroduction of the PtIP-50 polypeptide or a PtIP-65 polypeptidetranscript into the plant. Such methods include, for example,microinjection or particle bombardment. See, for example, Crossway, etal., (1986) Mol Gen. Genet. 202:179-185; Nomura, et al., (1986) PlantSci. 44:53-58; Hepler, et al., (1994) Proc. Natl. Acad. Sci.91:2176-2180 and Hush, et al., (1994) The Journal of Cell Science107:775-784, all of which are herein incorporated by reference.Alternatively, the PtIP-50 polypeptide and/or the PtIP-65 polypeptidepolynucleotide can be transiently transformed into the plant usingtechniques known in the art. Such techniques include viral vector systemand the precipitation of the polynucleotide in a manner that precludessubsequent release of the DNA. Thus, transcription from theparticle-bound DNA can occur, but the frequency with which it isreleased to become integrated into the genome is greatly reduced. Suchmethods include the use of particles coated with polyethylimine (PEI;Sigma #P3143).

Methods are known in the art for the targeted insertion of apolynucleotide at a specific location in the plant genome. In oneembodiment, the insertion of the polynucleotide at a desired genomiclocation is achieved using a site-specific recombination system. See,for example, WO 1999/25821, WO 1999/25854, WO 1999/25840, WO 1999/25855and WO 1999/25853, all of which are herein incorporated by reference.Briefly, the polynucleotide of the embodiments can be contained intransfer cassette flanked by two non-identical recombination sites. Thetransfer cassette is introduced into a plant have stably incorporatedinto its genome a target site which is flanked by two non-identicalrecombination sites that correspond to the sites of the transfercassette. An appropriate recombinase is provided and the transfercassette is integrated at the target site. The polynucleotide ofinterest is thereby integrated at a specific chromosomal position in theplant genome.

Plant transformation vectors may be comprised of one or more DNA vectorsneeded for achieving plant transformation. For example, it is a commonpractice in the art to utilize plant transformation vectors that arecomprised of more than one contiguous DNA segment. These vectors areoften referred to in the art as “binary vectors”. Binary vectors as wellas vectors with helper plasmids are most often used forAgrobacterium-mediated transformation, where the size and complexity ofDNA segments needed to achieve efficient transformation is quite large,and it is advantageous to separate functions onto separate DNAmolecules. Binary vectors typically contain a plasmid vector thatcontains the cis-acting sequences required for T-DNA transfer (such asleft border and right border), a selectable marker that is engineered tobe capable of expression in a plant cell, and a “gene of interest” (agene engineered to be capable of expression in a plant cell for whichgeneration of transgenic plants is desired). Also present on thisplasmid vector are sequences required for bacterial replication. Thecis-acting sequences are arranged in a fashion to allow efficienttransfer into plant cells and expression therein. For example, theselectable marker gene and the pesticidal gene are located between theleft and right borders. Often a second plasmid vector contains thetrans-acting factors that mediate T-DNA transfer from Agrobacterium toplant cells. This plasmid often contains the virulence functions (Virgenes) that allow infection of plant cells by Agrobacterium, andtransfer of DNA by cleavage at border sequences and vir-mediated DNAtransfer, as is understood in the art (Hellens and Mullineaux, (2000)Trends in Plant Science 5:446-451). Several types of Agrobacteriumstrains (e.g. LBA4404, GV3101, EHA101, EHA105, etc.) can be used forplant transformation. The second plasmid vector is not necessary fortransforming the plants by other methods such as microprojection,microinjection, electroporation, polyethylene glycol, etc.

In general, plant transformation methods involve transferringheterologous DNA into target plant cells (e.g., immature or matureembryos, suspension cultures, undifferentiated callus, protoplasts,etc.), followed by applying a maximum threshold level of appropriateselection (depending on the selectable marker gene) to recover thetransformed plant cells from a group of untransformed cell mass.Following integration of heterologous foreign DNA into plant cells, onethen applies a maximum threshold level of appropriate selection in themedium to kill the untransformed cells and separate and proliferate theputatively transformed cells that survive from this selection treatmentby transferring regularly to a fresh medium. By continuous passage andchallenge with appropriate selection, one identifies and proliferatesthe cells that are transformed with the plasmid vector. Molecular andbiochemical methods can then be used to confirm the presence of theintegrated heterologous gene of interest into the genome of thetransgenic plant.

Explants are typically transferred to a fresh supply of the same mediumand cultured routinely. Subsequently, the transformed cells aredifferentiated into shoots after placing on regeneration mediumsupplemented with a maximum threshold level of selecting agent. Theshoots are then transferred to a selective rooting medium for recoveringrooted shoot or plantlet. The transgenic plantlet then grows into amature plant and produces fertile seeds (e.g., Hiei, et al., (1994) ThePlant Journal 6:271-282; Ishida, et al., (1996) Nature Biotechnology14:745-750). Explants are typically transferred to a fresh supply of thesame medium and cultured routinely. A general description of thetechniques and methods for generating transgenic plants are found inAyres and Park, (1994) Critical Reviews in Plant Science 13:219-239 andBommineni and Jauhar, (1997) Maydica 42:107-120. Since the transformedmaterial contains many cells; both transformed and non-transformed cellsare present in any piece of subjected target callus or tissue or groupof cells. The ability to kill non-transformed cells and allowtransformed cells to proliferate results in transformed plant cultures.Often, the ability to remove non-transformed cells is a limitation torapid recovery of transformed plant cells and successful generation oftransgenic plants.

The cells that have been transformed may be grown into plants inaccordance with conventional ways. See, for example, McCormick, et al.,(1986) Plant Cell Reports 5:81-84. These plants may then be grown, andeither pollinated with the same transformed strain or different strains,and the resulting hybrid having constitutive or inducible expression ofthe desired phenotypic characteristic identified. Two or moregenerations may be grown to ensure that expression of the desiredphenotypic characteristic is stably maintained and inherited and thenseeds harvested to ensure that expression of the desired phenotypiccharacteristic has been achieved.

The nucleotide sequences of the embodiments may be provided to the plantby contacting the plant with a virus or viral nucleic acids. Generally,such methods involve incorporating the nucleotide construct of interestwithin a viral DNA or RNA molecule. It is recognized that therecombinant proteins of the embodiments may be initially synthesized aspart of a viral polyprotein, which later may be processed by proteolysisin vivo or in vitro to produce the desired PtIP-50 polypeptide or aPtIP-65 polypeptide. It is also recognized that such a viralpolyprotein, comprising at least a portion of the amino acid sequence ofa PtIP-50 polypeptide or a PtIP-65 polypeptide of the embodiments, mayhave the desired pesticidal activity. Such viral polyproteins and thenucleotide sequences that encode for them are encompassed by theembodiments. Methods for providing plants with nucleotide constructs andproducing the encoded proteins in the plants, which involve viral DNA orRNA molecules are known in the art. See, for example, U.S. Pat. Nos.5,889,191; 5,889,190; 5,866,785; 5,589,367 and 5,316,931; hereinincorporated by reference.

Methods for transformation of chloroplasts are known in the art. See,for example, Svab, et al., (1990) Proc. Natl. Acad. Sci. USA87:8526-8530; Svab and Maliga, (1993) Proc. Natl. Acad. Sci. USA90:913-917; Svab and Maliga, (1993) EMBO J. 12:601-606. The methodrelies on particle gun delivery of DNA containing a selectable markerand targeting of the DNA to the plastid genome through homologousrecombination. Additionally, plastid transformation can be accomplishedby transactivation of a silent plastid-borne transgene bytissue-preferred expression of a nuclear-encoded and plastid-directedRNA polymerase. Such a system has been reported in McBride, et al.,(1994) Proc. Natl. Acad. Sci. USA 91:7301-7305.

The embodiments further relate to plant-propagating material of atransformed plant of the embodiments including, but not limited to,seeds, tubers, corms, bulbs, leaves and cuttings of roots and shoots.

The embodiments may be used for transformation of any plant species,including, but not limited to, monocots and dicots. Examples of plantsof interest include, but are not limited to, corn (Zea mays), Brassicasp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassicaspecies useful as sources of seed oil, alfalfa (Medicago sativa), rice(Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghumvulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet(Panicum miliaceum), foxtail millet (Setaria italica), finger millet(Eleusine coracana)), sunflower (Helianthus annuus), safflower(Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycinemax), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts(Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum),sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee(Coffea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus),citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camelliasinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficuscasica), guava (Psidium guajava), mango (Mangifera indica), olive (Oleaeuropaea), papaya (Carica papaya), cashew (Anacardium occidentale),macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugarbeets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley,vegetables ornamentals, and conifers.

Vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g.,Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseoluslimensis), peas (Lathyrus spp.), and members of the genus Cucumis suchas cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon(C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea(Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosaspp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias(Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia(Euphorbia pulcherrima), and chrysanthemum. Conifers that may beemployed in practicing the embodiments include, for example, pines suchas loblolly pine (Pinus taeda), slash pine (Pinus ellioth), ponderosapine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Montereypine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Westernhemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood(Sequoia sempervirens); true firs such as silver fir (Abies amabilis)and balsam fir (Abies balsamea); and cedars such as Western red cedar(Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis).Plants of the embodiments include crop plants (for example, corn,alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut,sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants.

Turf grasses include, but are not limited to: annual bluegrass (Poaannua); annual ryegrass (Lolium multiflorum); Canada bluegrass (Poacompressa); Chewing's fescue (Festuca rubra); colonial bentgrass(Agrostis tenuis); creeping bentgrass (Agrostis palustris); crestedwheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyroncristatum); hard fescue (Festuca longifolia); Kentucky bluegrass (Poapratensis); orchardgrass (Dactylis glomerata); perennial ryegrass(Lolium perenne); red fescue (Festuca rubra); redtop (Agrostis alba);rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina); smoothbromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy(Phleum pratense); velvet bentgrass (Agrostis canina); weepingalkaligrass (Puccinellia distans); western wheatgrass (Agropyronsmithii); Bermuda grass (Cynodon spp.); St. Augustine grass(Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass(Paspalum notatum); carpet grass (Axonopus affinis); centipede grass(Eremochloa ophiuroides); kikuyu grass (Pennisetum clandesinum);seashore paspalum (Paspalum vaginatum); blue gramma (Boutelouagracilis); buffalo grass (Buchloe dactyloids); sideoats gramma(Bouteloua curtipendula).

Plants of interest include grain plants that provide seeds of interest,oil-seed plants, and leguminous plants. Seeds of interest include grainseeds, such as corn, wheat, barley, rice, sorghum, rye, millet, etc.Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica,maize, alfalfa, palm, coconut, flax, castor, olive, etc. Leguminousplants include beans and peas. Beans include guar, locust bean,fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, favabean, lentils, chickpea, etc.

Evaluation of Plant Transformation

Following introduction of heterologous foreign DNA into plant cells, thetransformation or integration of heterologous gene in the plant genomeis confirmed by various methods such as analysis of nucleic acids,proteins and metabolites associated with the integrated gene.

PCR analysis is a rapid method to screen transformed cells, tissue orshoots for the presence of incorporated gene at the earlier stage beforetransplanting into the soil (Sambrook and Russell, (2001) MolecularCloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y.). PCR is carried out using oligonucleotide primersspecific to the gene of interest or Agrobacterium vector background,etc.

Plant transformation may be confirmed by Southern blot analysis ofgenomic DNA (Sambrook and Russell, (2001) supra). In general, total DNAis extracted from the transformant, digested with appropriaterestriction enzymes, fractionated in an agarose gel and transferred to anitrocellulose or nylon membrane. The membrane or “blot” is then probedwith, for example, radiolabeled 32P target DNA fragment to confirm theintegration of introduced gene into the plant genome according tostandard techniques (Sambrook and Russell, (2001) supra).

In Northern blot analysis, RNA is isolated from specific tissues oftransformant, fractionated in a formaldehyde agarose gel, and blottedonto a nylon filter according to standard procedures that are routinelyused in the art (Sambrook and Russell, (2001) supra). Expression of RNAencoded by the pesticidal gene is then tested by hybridizing the filterto a radioactive probe derived from a pesticidal gene, by methods knownin the art (Sambrook and Russell, (2001) supra).

Western blot, biochemical assays and the like may be carried out on thetransgenic plants to confirm the presence of protein encoded by thepesticidal gene by standard procedures (Sambrook and Russell, 2001,supra) using antibodies that bind to one or more epitopes present on thePtIP-50 polypeptide or the PtIP-65 polypeptide.

Stacking of Traits in Transgenic Plant

Transgenic plants may comprise a stack of one or more insecticidalpolynucleotides disclosed herein with one or more additionalpolynucleotides resulting in the production or suppression of multiplepolypeptide sequences. Transgenic plants comprising stacks ofpolynucleotide sequences can be obtained by either or both oftraditional breeding methods or through genetic engineering methods.These methods include, but are not limited to, breeding individual lineseach comprising a polynucleotide of interest, transforming a transgenicplant comprising a gene disclosed herein with a subsequent gene andco-transformation of genes into a single plant cell. As used herein, theterm “stacked” includes having the multiple traits present in the sameplant (i.e., both traits are incorporated into the nuclear genome, onetrait is incorporated into the nuclear genome and one trait isincorporated into the genome of a plastid or both traits areincorporated into the genome of a plastid). In one non-limiting example,“stacked traits” comprise a molecular stack where the sequences arephysically adjacent to each other. A trait, as used herein, refers tothe phenotype derived from a particular sequence or groups of sequences.Co-transformation of genes can be carried out using singletransformation vectors comprising multiple genes or genes carriedseparately on multiple vectors. If the sequences are stacked bygenetically transforming the plants, the polynucleotide sequences ofinterest can be combined at any time and in any order. The traits can beintroduced simultaneously in a co-transformation protocol with thepolynucleotides of interest provided by any combination oftransformation cassettes. For example, if two sequences will beintroduced, the two sequences can be contained in separatetransformation cassettes (trans) or contained on the same transformationcassette (cis). Expression of the sequences can be driven by the samepromoter or by different promoters. In certain cases, it may bedesirable to introduce a transformation cassette that will suppress theexpression of the polynucleotide of interest. This may be combined withany combination of other suppression cassettes or overexpressioncassettes to generate the desired combination of traits in the plant. Itis further recognized that polynucleotide sequences can be stacked at adesired genomic location using a site-specific recombination system.See, for example, WO 1999/25821, WO 1999/25854, WO 1999/25840, WO1999/25855 and WO 1999/25853, all of which are herein incorporated byreference.

In some embodiments the polynucleotides encoding the PtIP-50 polypeptideor the PtIP-65 polypeptide disclosed herein, alone or stacked with oneor more additional insect resistance traits can be stacked with one ormore additional input traits (e.g., herbicide resistance, fungalresistance, virus resistance, stress tolerance, disease resistance, malesterility, stalk strength, and the like) or output traits (e.g.,increased yield, modified starches, improved oil profile, balanced aminoacids, high lysine or methionine, increased digestibility, improvedfiber quality, drought resistance, and the like). Thus, thepolynucleotide embodiments can be used to provide a complete agronomicpackage of improved crop quality with the ability to flexibly and costeffectively control any number of agronomic pests.

Transgenes useful for stacking include but are not limited to:

1. Transgenes that Confer Resistance to Insects or Disease and thatEncode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example, Jones, et al., (1994) Science266:789 (cloning of the tomato Cf-9 gene for resistance to Cladosporiumfulvum); Martin, et al., (1993) Science 262:1432 (tomato Pto gene forresistance to Pseudomonas syringae pv. tomato encodes a protein kinase);Mindrinos, et al., (1994) Cell 78:1089 (Arabidopsis RSP2 gene forresistance to Pseudomonas syringae), McDowell and Woffenden, (2003)Trends Biotechnol. 21(4):178-83 and Toyoda, et al., (2002) TransgenicRes. 11(6):567-82. A plant resistant to a disease is one that is moreresistant to a pathogen as compared to the wild type plant.

(B) Genes encoding a Bacillus thuringiensis protein, a derivativethereof or a synthetic polypeptide modeled thereon. See, for example,Geiser, et al., (1986) Gene 48:109, who disclose the cloning andnucleotide sequence of a Bt delta-endotoxin gene. Moreover, DNAmolecules encoding delta-endotoxin genes can be purchased from AmericanType Culture Collection (Rockville, Md.), for example, under ATCC®Accession Numbers 40098, 67136, 31995 and 31998. Other non-limitingexamples of Bacillus thuringiensis transgenes being geneticallyengineered are given in the following patents and patent applicationsand hereby are incorporated by reference for this purpose: U.S. Pat.Nos. 5,188,960; 5,689,052; 5,880,275; 5,986,177; 6,023,013, 6,060,594,6,063,597, 6,077,824, 6,620,988, 6,642,030, 6,713,259, 6,893,826,7,105,332; 7,179,965, 7,208,474; 7,227,056, 7,288,643, 7,323,556,7,329,736, 7,449,552, 7,468,278, 7,510,878, 7,521,235, 7,544,862,7,605,304, 7,696,412, 7,629,504, 7,705,216, 7,772,465, 7,790,846,7,858,849 and WO 1991/14778; WO 1999/31248; WO 2001/12731; WO 1999/24581and WO 1997/40162.

Genes encoding pesticidal proteins may also be stacked including but arenot limited to: insecticidal proteins from Pseudomonas sp. such asPSEEN3174 (Monalysin, (2011) PLoS Pathogens, 7:1-13), from Pseudomonasprotegens strain CHAO and Pf-5 (previously fluorescens) (Pechy-Tarr,(2008) Environmental Microbiology 10:2368-2386: GenBank Accession No.EU400157); from Pseudomonas Taiwanensis (Liu, et al., (2010) J. Agric.Food Chem. 58:12343-12349) and from Pseudomonas pseudoalcligenes (Zhang,et al., (2009) Annals of Microbiology 59:45-50 and Li, et al., (2007)Plant Cell Tiss. Organ Cult. 89:159-168); insecticidal proteins fromPhotorhabdus sp. and Xenorhabdus sp. (Hinchliffe, et al., (2010) TheOpen Toxinology Journal 3:101-118 and Morgan, et al., (2001) Applied andEnvir. Micro. 67:2062-2069), U.S. Pat. Nos. 6,048,838, and 6,379,946; aPIP-1 polypeptide of U.S. Ser. No. 13/792,861; an AfIP-1A and/or AfIP-1Bpolypeptide of U.S. Ser. No. 13/800,233; a PHI-4 polypeptide of U.S.Ser. No. 13/839,702; and δ-endotoxins including, but not limited to, theCry1, Cry2, Cry3, Cry4, Cry5, Cry6, Cry7, Cry8, Cry9, Cry10, Cry1I,Cry12, Cry13, Cry14, Cry15, Cry16, Cry17, Cry18, Cry19, Cry20, Cry21,Cry22, Cry23, Cry24, Cry25, Cry26, Cry27, Cry 28, Cry 29, Cry 30, Cry31,Cry32, Cry33, Cry34, Cry35, Cry36, Cry37, Cry38, Cry39, Cry40, Cry41,Cry42, Cry43, Cry44, Cry45, Cry 46, Cry47, Cry49, Cry 51 and Cry55classes of 5-endotoxin genes and the B. thuringiensis cytolytic Cyt1 andCyt2 genes. Members of these classes of B. thuringiensis insecticidalproteins include, but are not limited to Cry1Aa1 (Accession # AAA22353);Cry1Aa2 (Accession # Accession # AAA22552); Cry1Aa3 (Accession #BAA00257); Cry1Aa4 (Accession # CAA31886); Cry1Aa5 (Accession #BAA04468); Cry1Aa6 (Accession # AAA86265); Cry1Aa7 (Accession #AAD46139); Cry1Aa8 (Accession # I26149); Cry1Aa9 (Accession # BAA77213);Cry1Aa10 (Accession # AAD55382); Cry1Aa11 (Accession # CAA70856);Cry1Aa12 (Accession # AAP80146); Cry1Aa13 (Accession # AAM44305);Cry1Aa14 (Accession # AAP40639); Cry1Aa15 (Accession # AAY66993);Cry1Aa16 (Accession # HQ439776); Cry1Aa17 (Accession # HQ439788);Cry1Aa18 (Accession # HQ439790); Cry1Aa19 (Accession # HQ685121);Cry1Aa20 (Accession # JF340156); Cry1Aa21 (Accession # JN651496);Cry1Aa22 (Accession # KC158223); Cry1Ab1 (Accession # AAA22330); Cry1Ab2(Accession # AAA22613); Cry1Ab3 (Accession # AAA22561); Cry1Ab4(Accession # BAA00071); Cry1Ab5 (Accession # CAA28405); Cry1Ab6(Accession # AAA22420); Cry1Ab7 (Accession # CAA31620); Cry1Ab8(Accession # AAA22551); Cry1Ab9 (Accession # CAA38701); Cry1Ab10(Accession # A29125); Cry1Ab11 (Accession # I12419); Cry1Ab12 (Accession# AAC64003); Cry1Ab13 (Accession # AAN76494); Cry1Ab14 (Accession #AAG16877); Cry1Ab15 (Accession # AAO13302); Cry1Ab16 (Accession #AAK55546); Cry1Ab17 (Accession # AAT46415); Cry1Ab18 (Accession #AAQ88259); Cry1Ab19 (Accession # AAW31761); Cry1Ab20 (Accession #ABB72460); Cry1Ab21 (Accession # ABS18384); Cry1Ab22 (Accession #ABW87320); Cry1Ab23 (Accession # HQ439777); Cry1Ab24 (Accession #HQ439778); Cry1Ab25 (Accession # HQ685122); Cry1Ab26 (Accession #HQ847729); Cry1Ab27 (Accession # JN135249); Cry1Ab28 (Accession #JN135250); Cry1Ab29 (Accession # JN135251); Cry1Ab30 (Accession #JN135252); Cry1Ab31 (Accession # JN135253); Cry1Ab32 (Accession #JN135254); Cry1Ab33 (Accession # AAS93798); Cry1Ab34 (Accession #KC156668); Cry1Ab-like (Accession # AAK14336); Cry1Ab-like (Accession #AAK14337); Cry1Ab-like (Accession # AAK14338); Cry1Ab-like (Accession #ABG88858); Cry1Ac1 (Accession # AAA22331); Cry1Ac2 (Accession #AAA22338); Cry1Ac3 (Accession # CAA38098); Cry1Ac4 (Accession #AAA73077); Cry1Ac5 (Accession # AAA22339); Cry1Ac6 (Accession #AAA86266); Cry1Ac7 (Accession # AAB46989); Cry1Ac8 (Accession #AAC44841); Cry1Ac9 (Accession # AAB49768); Cry1Ac10 (Accession #CAA05505); Cry1Ac11 (Accession # CAA10270); Cry1Ac12 (Accession #I12418); Cry1Ac13 (Accession # AAD38701); Cry1Ac14 (Accession #AAQ06607); Cry1Ac15 (Accession # AAN07788); Cry1Ac16 (Accession #AAU87037); Cry1Ac17 (Accession # AAX18704); Cry1Ac18 (Accession #AAY88347); Cry1Ac19 (Accession # ABD37053); Cry1Ac20 (Accession #ABB89046); Cry1Ac21 (Accession # AAY66992); Cry1Ac22 (Accession #ABZ01836); Cry1Ac23 (Accession # CAQ30431); Cry1Ac24 (Accession #ABL01535); Cry1Ac25 (Accession # FJ513324); Cry1Ac26 (Accession #FJ617446); Cry1Ac27 (Accession # FJ617447); Cry1Ac28 (Accession #ACM90319); Cry1Ac29 (Accession # DQ438941); Cry1Ac30 (Accession #GQ227507); Cry1Ac31 (Accession # GU446674); Cry1Ac32 (Accession #HM061081); Cry1Ac33 (Accession # GQ866913); Cry1Ac34 (Accession #HQ230364); Cry1Ac35 (Accession # JF340157); Cry1Ac36 (Accession #JN387137); Cry1Ac37 (Accession # JQ317685); Cry1Ad1 (Accession #AAA22340); Cry1Ad2 (Accession # CAA01880); Cry1Ae1 (Accession #AAA22410); Cry1Af1 (Accession # AAB82749); Cry1Ag1 (Accession #AAD46137); Cry1Ah1 (Accession # AAQ14326); Cry1Ah2 (Accession #ABB76664); Cry1Ah3 (Accession # HQ439779); Cry1Ai1 (Accession #AAO39719); Cry1Ai2 (Accession # HQ439780); Cry1A-like (Accession #AAK14339); Cry1Ba1 (Accession # CAA29898); Cry1Ba2 (Accession #CAA65003); Cry1Ba3 (Accession # AAK63251); Cry1Ba4 (Accession #AAK51084); Cry1Ba5 (Accession # ABO20894); Cry1Ba6 (Accession #ABL60921); Cry1Ba7 (Accession # HQ439781); Cry1Bb1 (Accession #AAA22344); Cry1Bb2 (Accession # HQ439782); Cry1Bc1 (Accession #CAA86568); Cry1Bd1 (Accession # AAD10292); Cry1Bd2 (Accession #AAM93496); Cry1Be1 (Accession # AAC32850); Cry1Be2 (Accession #AAQ52387); Cry1Be3 (Accession # ACV96720); Cry1Be4 (Accession #HM070026); Cry1Bf1 (Accession # CAC50778); Cry1Bf2 (Accession #AAQ52380); Cry1Bg1 (Accession # AAO39720); Cry1Bh1 (Accession #HQ589331); Cry1Bi1 (Accession # KC156700); Cry1Ca1 (Accession #CAA30396); Cry1Ca2 (Accession # CAA31951); Cry1Ca3 (Accession #AAA22343); Cry1Ca4 (Accession # CAA01886); Cry1Ca5 (Accession #CAA65457); Cry1Ca6 [1] (Accession # AAF37224); Cry1Ca7 (Accession #AAG50438); Cry1Ca8 (Accession # AAM00264); Cry1Ca9 (Accession #AAL79362); Cry1Ca10 (Accession # AAN16462); Cry1Ca11 (Accession #AAX53094); Cry1Ca12 (Accession # HM070027); Cry1Ca13 (Accession #HQ412621); Cry1Ca14 (Accession # JN651493); Cry1Cb1 (Accession #M97880); Cry1Cb2 (Accession # AAG35409); Cry1Cb3 (Accession # ACD50894);Cry1Cb-like (Accession # AAX63901); Cry1Da1 (Accession # CAA38099);Cry1Da2 (Accession # I76415); Cry1Da3 (Accession # HQ439784); Cry1Db1(Accession # CAA80234); Cry1Db2 (Accession # AAK48937); Cry1Dc1(Accession # ABK35074); Cry1Ea1 (Accession # CAA37933); Cry1Ea2(Accession # CAA39609); Cry1Ea3 (Accession # AAA22345); Cry1Ea4(Accession # AAD04732); Cry1Ea5 (Accession # A15535); Cry1Ea6 (Accession# AAL50330); Cry1Ea7 (Accession # AAW72936); Cry1Ea8 (Accession #ABX11258); Cry1Ea9 (Accession # HQ439785); Cry1Ea10 (Accession #ADR00398); Cry1Ea11 (Accession # JQ652456); Cry1Eb1 (Accession #AAA22346); Cry1Fa1 (Accession # AAA22348); Cry1Fa2 (Accession #AAA22347); Cry1Fa3 (Accession # HM070028); Cry1Fa4 (Accession #HM439638); Cry1Fb1 (Accession # CAA80235); Cry1Fb2 (Accession #BAA25298); Cry1Fb3 (Accession # AAF21767); Cry1Fb4 (Accession #AAC10641); Cry1Fb5 (Accession # AAO13295); Cry1Fb6 (Accession #ACD50892); Cry1Fb7 (Accession # ACD50893); Cry1Ga1 (Accession #CAA80233); Cry1Ga2 (Accession # CAA70506); Cry1Gb1 (Accession #AAD10291); Cry1Gb2 (Accession # AAO13756); Cry1Gc1 (Accession #AAQ52381); Cry1Ha1 (Accession # CAA80236); Cry1Hb1 (Accession #AAA79694); Cry1Hb2 (Accession # HQ439786); Cry1H-like (Accession #AAF01213); Cry1Ia1 (Accession # CAA44633); Cry1Ia2 (Accession #AAA22354); Cry1Ia3 (Accession # AAC36999); Cry1Ia4 (Accession #AAB00958); Cry1Ia5 (Accession # CAA70124); Cry1Ia6 (Accession #AAC26910); Cry1Ia7 (Accession # AAM73516); Cry1Ia8 (Accession #AAK66742); Cry1Ia9 (Accession # AAQ08616); Cry1Ia10 (Accession #AAP86782); Cry1Ia11 (Accession # CAC85964); Cry1Ia12 (Accession #AAV53390); Cry1Ia13 (Accession # ABF83202); Cry1Ia14 (Accession #ACG63871); Cry1Ia15 (Accession # FJ617445); Cry1Ia16 (Accession #FJ617448); Cry1Ia17 (Accession # GU989199); Cry1Ia18 (Accession #ADK23801); Cry1Ia19 (Accession # HQ439787); Cry1Ia20 (Accession #JQ228426); Cry1Ia21 (Accession # JQ228424); Cry1Ia22 (Accession #JQ228427); Cry1Ia23 (Accession # JQ228428); Cry1Ia24 (Accession #JQ228429); Cry1Ia25 (Accession # JQ228430); Cry1Ia26 (Accession #JQ228431); Cry1Ia27 (Accession # JQ228432); Cry1Ia28 (Accession #JQ228433); Cry1Ia29 (Accession # JQ228434); Cry1Ia30 (Accession #JQ317686); Cry1Ia31 (Accession # JX944038); Cry1Ia32 (Accession #JX944039); Cry1Ia33 (Accession # JX944040); Cry1Ib1 (Accession #AAA82114); Cry1Ib2 (Accession # ABW88019); Cry1Ib3 (Accession #ACD75515); Cry1Ib4 (Accession # HM051227); Cry1Ib5 (Accession #HM070028); Cry1Ib6 (Accession # ADK38579); Cry1Ib7 (Accession #JN571740); Cry1Ib8 (Accession # JN675714); Cry1Ib9 (Accession #JN675715); Cry1Ib10 (Accession # JN675716); Cry1Ib11 (Accession #JQ228423); Cry1Ic1 (Accession # AAC62933); Cry1Ic2 (Accession #AAE71691); Cry1Id1 (Accession # AAD44366); Cry1Id2 (Accession #JQ228422); Cry1Ie1 (Accession # AAG43526); Cry1Ie2 (Accession #HM439636); Cry1Ie3 (Accession # KC156647); Cry1Ie4 (Accession #KC156681); Cry1If1 (Accession # AAQ52382); Cry1Ig1 (Accession #KC156701); Cry1I-like (Accession # AAC31094); Cry1I-like (Accession #ABG88859); Cry1Ja1 (Accession # AAA22341); Cry1Ja2 (Accession #HM070030); Cry1Ja3 (Accession # JQ228425); Cry1Jb1 (Accession #AAA98959); Cry1Jc1 (Accession # AAC31092); Cry1Jc2 (Accession #AAQ52372); Cry1Jd1 (Accession # CAC50779); Cry1Ka1 (Accession #AAB00376); Cry1Ka2 (Accession # HQ439783); Cry1La1 (Accession #AAS60191); Cry1La2 (Accession # HM070031); Cry1Ma1 (Accession #FJ884067); Cry1Ma2 (Accession # KC156659); Cry1Na1 (Accession #KC156648); Cry1Nb1 (Accession # KC156678); Cry1-like (Accession #AAC31091); Cry2Aa1 (Accession # AAA22335); Cry2Aa2 (Accession #AAA83516); Cry2Aa3 (Accession # D86064); Cry2Aa4 (Accession # AAC04867);Cry2Aa5 (Accession # CAA10671); Cry2Aa6 (Accession # CAA10672); Cry2Aa7(Accession # CAA10670); Cry2Aa8 (Accession # AAO13734); Cry2Aa9(Accession # AAO13750); Cry2Aa10 (Accession # AAQ04263); Cry2Aa11(Accession # AAQ52384); Cry2Aa12 (Accession # ABI83671); Cry2Aa13(Accession # ABL01536); Cry2Aa14 (Accession # ACF04939); Cry2Aa15(Accession # JN426947); Cry2Ab1 (Accession # AAA22342); Cry2Ab2(Accession # CAA39075); Cry2Ab3 (Accession # AAG36762); Cry2Ab4(Accession # AAO13296); Cry2Ab5 (Accession # AAQ04609); Cry2Ab6(Accession # AAP59457); Cry2Ab7 (Accession # AAZ66347); Cry2Ab8(Accession # ABC95996); Cry2Ab9 (Accession # ABC74968); Cry2Ab10(Accession # EF157306); Cry2Ab11 (Accession # CAM84575); Cry2Ab12(Accession # ABM21764); Cry2Ab13 (Accession # ACG76120); Cry2Ab14(Accession # ACG76121); Cry2Ab15 (Accession # HM037126); Cry2Ab16(Accession # GQ866914); Cry2Ab17 (Accession # HQ439789); Cry2Ab18(Accession # JN135255); Cry2Ab19 (Accession # JN135256); Cry2Ab20(Accession # JN135257); Cry2Ab21 (Accession # JN135258); Cry2Ab22(Accession # JN135259); Cry2Ab23 (Accession # JN135260); Cry2Ab24(Accession # JN135261); Cry2Ab25 (Accession # JN415485); Cry2Ab26(Accession # JN426946); Cry2Ab27 (Accession # JN415764); Cry2Ab28(Accession # JN651494); Cry2Ac1 (Accession # CAA40536); Cry2Ac2(Accession # AAG35410); Cry2Ac3 (Accession # AAQ52385); Cry2Ac4(Accession # ABC95997); Cry2Ac5 (Accession # ABC74969); Cry2Ac6(Accession # ABC74793); Cry2Ac7 (Accession # CAL18690); Cry2Ac8(Accession # CAM09325); Cry2Ac9 (Accession # CAM09326); Cry2Ac10(Accession # ABN15104); Cry2Ac11 (Accession # CAM83895); Cry2Ac12(Accession # CAM83896); Cry2Ad1 (Accession # AAF09583); Cry2Ad2(Accession # ABC86927); Cry2Ad3 (Accession # CAK29504); Cry2Ad4(Accession # CAM32331); Cry2Ad5 (Accession # CAO78739); Cry2Ae1(Accession # AAQ52362); Cry2Af1 (Accession # ABO30519); Cry2Af2(Accession # GQ866915); Cry2Ag1 (Accession # ACH91610); Cry2Ah1(Accession # EU939453); Cry2Ah2 (Accession # ACL80665); Cry2Ah3(Accession # GU073380); Cry2Ah4 (Accession # KC156702); Cry2Ai1(Accession # FJ788388); Cry2Aj (Accession #); Cry2Ak1 (Accession #KC156660); Cry2Ba1 (Accession # KC156658); Cry3Aa1 (Accession #AAA22336); Cry3Aa2 (Accession # AAA22541); Cry3Aa3 (Accession #CAA68482); Cry3Aa4 (Accession # AAA22542); Cry3Aa5 (Accession #AAA50255); Cry3Aa6 (Accession # AAC43266); Cry3Aa7 (Accession #CAB41411); Cry3Aa8 (Accession # AAS79487); Cry3Aa9 (Accession #AAW05659); Cry3Aa10 (Accession # AAU29411); Cry3Aa11 (Accession #AAW82872); Cry3Aa12 (Accession # ABY49136); Cry3Ba1 (Accession #CAA34983); Cry3Ba2 (Accession # CAA00645); Cry3Ba3 (Accession #JQ397327); Cry3Bb1 (Accession # AAA22334); Cry3Bb2 (Accession #AAA74198); Cry3Bb3 (Accession # I15475); Cry3Ca1 (Accession # CAA42469);Cry4Aa1 (Accession # CAA68485); Cry4Aa2 (Accession # BAA00179); Cry4Aa3(Accession # CAD30148); Cry4Aa4 (Accession # AFB18317); Cry4A-like(Accession # AAY96321); Cry4Ba1 (Accession # CAA30312); Cry4Ba2(Accession # CAA30114); Cry4Ba3 (Accession # AAA22337); Cry4Ba4(Accession # BAA00178); Cry4Ba5 (Accession # CAD30095); Cry4Ba-like(Accession # ABC47686); Cry4Ca1 (Accession # EU646202); Cry4Cb1(Accession # FJ403208); Cry4Cb2 (Accession # FJ597622); Cry4Cc1(Accession # FJ403207); Cry5Aa1 (Accession # AAA67694); Cry5Ab1(Accession # AAA67693); Cry5Ac1 (Accession # I34543); Cry5Ad1 (Accession# ABQ82087); Cry5Ba1 (Accession # AAA68598); Cry5Ba2 (Accession #ABW88931); Cry5Ba3 (Accession # AFJ04417); Cry5Ca1 (Accession #HM461869); Cry5Ca2 (Accession # ZP_04123426); Cry5Da1 (Accession #HM461870); Cry5Da2 (Accession # ZP_04123980); Cry5Ea1 (Accession #HM485580); Cry5Ea2 (Accession # ZP_04124038); Cry6Aa1 (Accession #AAA22357); Cry6Aa2 (Accession # AAM46849); Cry6Aa3 (Accession #ABH03377); Cry6Ba1 (Accession # AAA22358); Cry7Aa1 (Accession #AAA22351); Cry7Ab1 (Accession # AAA21120); Cry7Ab2 (Accession #AAA21121); Cry7Ab3 (Accession # ABX24522); Cry7Ab4 (Accession #EU380678); Cry7Ab5 (Accession # ABX79555); Cry7Ab6 (Accession #ACI44005); Cry7Ab7 (Accession # ADB89216); Cry7Ab8 (Accession #GU145299); Cry7Ab9 (Accession # ADD92572); Cry7Ba1 (Accession #ABB70817); Cry7Bb1 (Accession # KC156653); Cry7Ca1 (Accession #ABR67863); Cry7Cb1 (Accession # KC156698); Cry7Da1 (Accession #ACQ99547); Cry7Da2 (Accession # HM572236); Cry7Da3 (Accession #KC156679); Cry7Ea1 (Accession # HM035086); Cry7Ea2 (Accession #HM132124); Cry7Ea3 (Accession # EEM19403); Cry7Fa1 (Accession #HM035088); Cry7Fa2 (Accession # EEM19090); Cry7Fb1 (Accession #HM572235); Cry7Fb2 (Accession # KC156682); Cry7Ga1 (Accession #HM572237); Cry7Ga2 (Accession # KC156669); Cry7Gb1 (Accession #KC156650); Cry7Gc1 (Accession # KC156654); Cry7Gd1 (Accession #KC156697); Cry7Ha1 (Accession # KC156651); Cry7Ia1 (Accession #KC156665); Cry7Ja1 (Accession # KC156671); Cry7Ka1 (Accession #KC156680); Cry7Kb1 (Accession # BAM99306); Cry7La1 (Accession #BAM99307); Cry8Aa1 (Accession # AAA21117); Cry8Ab1 (Accession #EU044830); Cry8Ac1 (Accession # KC156662); Cry8Ad1 (Accession #KC156684); Cry8Ba1 (Accession # AAA21118); Cry8Bb1 (Accession #CAD57542); Cry8Bc1 (Accession # CAD57543); Cry8Ca1 (Accession #AAA21119); Cry8Ca2 (Accession # AAR98783); Cry8Ca3 (Accession #EU625349); Cry8Ca4 (Accession # ADB54826); Cry8Da1 (Accession #BAC07226); Cry8Da2 (Accession # BD133574); Cry8Da3 (Accession #BD133575); Cry8Db1 (Accession # BAF93483); Cry8Ea1 (Accession #AAQ73470); Cry8Ea2 (Accession # EU047597); Cry8Ea3 (Accession #KC855216); Cry8Fa1 (Accession # AAT48690); Cry8Fa2 (Accession #HQ174208); Cry8Fa3 (Accession # AFH78109); Cry8Ga1 (Accession #AAT46073); Cry8Ga2 (Accession # ABC42043); Cry8Ga3 (Accession #FJ198072); Cry8Ha1 (Accession # AAW81032); Cry8Ia1 (Accession #EU381044); Cry81a2 (Accession # GU073381); Cry81a3 (Accession #HM044664); Cry81a4 (Accession # KC156674); Cry8Ib1 (Accession #GU325772); Cry81b2 (Accession # KC156677); Cry8Ja1 (Accession #EU625348); Cry8Ka1 (Accession # FJ422558); Cry8Ka2 (Accession #ACN87262); Cry8Kb1 (Accession # HM123758); Cry8Kb2 (Accession #KC156675); Cry8La1 (Accession # GU325771); Cry8Ma1 (Accession #HM044665); Cry8Ma2 (Accession # EEM86551); Cry8Ma3 (Accession #HM210574); Cry8Na1 (Accession # HM640939); Cry8Pa1 (Accession #HQ388415); Cry8Qa1 (Accession # HQ441166); Cry8Qa2 (Accession #KC152468); Cry8Ra1 (Accession # AFP87548); Cry8Sa1 (Accession #JQ740599); Cry8Ta1 (Accession # KC156673); Cry8-like (Accession #FJ770571); Cry8-like (Accession # ABS53003); Cry9Aa1 (Accession #CAA41122); Cry9Aa2 (Accession # CAA41425); Cry9Aa3 (Accession #GQ249293); Cry9Aa4 (Accession # GQ249294); Cry9Aa5 (Accession #JX174110); Cry9Aa like (Accession # AAQ52376); Cry9Ba1 (Accession #CAA52927); Cry9Ba2 (Accession # GU299522); Cry9Bb1 (Accession #AAV28716); Cry9Ca1 (Accession # CAA85764); Cry9Ca2 (Accession #AAQ52375); Cry9Da1 (Accession # BAA19948); Cry9Da2 (Accession #AAB97923); Cry9Da3 (Accession # GQ249293); Cry9Da4 (Accession #GQ249297); Cry9Db1 (Accession # AAX78439); Cry9Dc1 (Accession #KC156683); Cry9Ea1 (Accession # BAA34908); Cry9Ea2 (Accession #AAO12908); Cry9Ea3 (Accession # ABM21765); Cry9Ea4 (Accession #ACE88267); Cry9Ea5 (Accession # ACF04743); Cry9Ea6 (Accession #ACG63872); Cry9Ea7 (Accession # FJ380927); Cry9Ea8 (Accession #GQ249292); Cry9Ea9 (Accession # JN651495); Cry9Eb1 (Accession #CAC50780); Cry9Eb2 (Accession # GQ249298); Cry9Eb3 (Accession #KC156646); Cry9Ec1 (Accession # AAC63366); Cry9Ed1 (Accession #AAX78440); Cry9Ee1 (Accession # GQ249296); Cry9Ee2 (Accession #KC156664); Cry9Fa1 (Accession # KC156692); Cry9Ga1 (Accession #KC156699); Cry9-like (Accession # AAC63366); Cry10Aa1 (Accession #AAA22614); Cry10Aa2 (Accession # E00614); Cry10Aa3 (Accession #CAD30098); Cry10Aa4 (Accession # AFB18318); Cry10A-like (Accession #DQ167578); Cry11Aa1 (Accession # AAA22352); Cry11Aa2 (Accession #AAA22611); Cry11Aa3 (Accession # CAD30081); Cry11Aa4 (Accession #AFB18319); Cry11Aa-like (Accession # DQ166531); Cry11Ba1 (Accession #CAA60504); Cry11Bb1 (Accession # AAC97162); Cry11Bb2 (Accession #HM068615); Cry12Aa1 (Accession # AAA22355); Cry13Aa1 (Accession #AAA22356); Cry14Aa1 (Accession # AAA21516); Cry14Ab1 (Accession #KC156652); Cry15Aa1 (Accession # AAA22333); Cry16Aa1 (Accession #CAA63860); Cry17Aa1 (Accession # CAA67841); Cry18Aa1 (Accession #CAA67506); Cry18Ba1 (Accession # AAF89667); Cry18Ca1 (Accession #AAF89668); Cry19Aa1 (Accession # CAA68875); Cry19Ba1 (Accession #BAA32397); Cry19Ca1 (Accession # AFM37572); Cry20Aa1 (Accession #AAB93476); Cry20Ba1 (Accession # ACS93601); Cry20Ba2 (Accession #KC156694); Cry20-like (Accession # GQ144333); Cry21Aa1 (Accession #I32932); Cry21Aa2 (Accession # I66477); Cry21Ba1 (Accession # BAC06484);Cry21Ca1 (Accession # JF521577); Cry21Ca2 (Accession # KC156687);Cry21Da1 (Accession # JF521578); Cry22Aa1 (Accession # I34547); Cry22Aa2(Accession # CAD43579); Cry22Aa3 (Accession # ACD93211); Cry22Ab1(Accession # AAK50456); Cry22Ab2 (Accession # CAD43577); Cry22Ba1(Accession # CAD43578); Cry22Bb1 (Accession # KC156672); Cry23Aa1(Accession # AAF76375); Cry24Aa1 (Accession # AAC61891); Cry24Ba1(Accession # BAD32657); Cry24Ca1 (Accession # CAJ43600); Cry25Aa1(Accession # AAC61892); Cry26Aa1 (Accession # AAD25075); Cry27Aa1(Accession # BAA82796); Cry28Aa1 (Accession # AAD24189); Cry28Aa2(Accession # AAG00235); Cry29Aa1 (Accession # CAC80985); Cry30Aa1(Accession # CAC80986); Cry30Ba1 (Accession # BAD00052); Cry30Ca1(Accession # BAD67157); Cry30Ca2 (Accession # ACU24781); Cry30Da1(Accession # EF095955); Cry30Db1 (Accession # BAE80088); Cry30Ea1(Accession # ACC95445); Cry30Ea2 (Accession # FJ499389); Cry30Fa1(Accession # ACI22625); Cry30Ga1 (Accession # ACG60020); Cry30Ga2(Accession # HQ638217); Cry31Aa1 (Accession # BAB11757); Cry31Aa2(Accession # AAL87458); Cry31Aa3 (Accession # BAE79808); Cry31Aa4(Accession # BAF32571); Cry31Aa5 (Accession # BAF32572); Cry31Aa6(Accession # BA144026); Cry31Ab1 (Accession # BAE79809); Cry31Ab2(Accession # BAF32570); Cry31Ac1 (Accession # BAF34368); Cry31Ac2(Accession # AB731600); Cry31Ad1 (Accession # BA144022); Cry32Aa1(Accession # AAG36711); Cry32Aa2 (Accession # GU063849); Cry32Ab1(Accession # GU063850); Cry32Ba1 (Accession # BAB78601); Cry32Ca1(Accession # BAB78602); Cry32Cb1 (Accession # KC156708); Cry32Da1(Accession # BAB78603); Cry32Ea1 (Accession # GU324274); Cry32Ea2(Accession # KC156686); Cry32Eb1 (Accession # KC156663); Cry32Fa1(Accession # KC156656); Cry32Ga1 (Accession # KC156657); Cry32Ha1(Accession # KC156661); Cry32Hb1 (Accession # KC156666); Cry321a1(Accession # KC156667); Cry32Ja1 (Accession # KC156685); Cry32Ka1(Accession # KC156688); Cry32La1 (Accession # KC156689); Cry32Ma1(Accession # KC156690); Cry32Mb1 (Accession # KC156704); Cry32Na1(Accession # KC156691); Cry32Oa1 (Accession # KC156703); Cry32Pa1(Accession # KC156705); Cry32Qa1 (Accession # KC156706); Cry32Ra1(Accession # KC156707); Cry32Sa1 (Accession # KC156709); Cry32Ta1(Accession # KC156710); Cry32Ua1 (Accession # KC156655); Cry33Aa1(Accession # AAL26871); Cry34Aa1 (Accession # AAG50341); Cry34Aa2(Accession # AAK64560); Cry34Aa3 (Accession # AAT29032); Cry34Aa4(Accession # AAT29030); Cry34Ab1 (Accession # AAG41671); Cry34Ac1(Accession # AAG50118); Cry34Ac2 (Accession # AAK64562); Cry34Ac3(Accession # AAT29029); Cry34Ba1 (Accession # AAK64565); Cry34Ba2(Accession # AAT29033); Cry34Ba3 (Accession # AAT29031); Cry35Aa1(Accession # AAG50342); Cry35Aa2 (Accession # AAK64561); Cry35Aa3(Accession # AAT29028); Cry35Aa4 (Accession # AAT29025); Cry35Ab1(Accession # AAG41672); Cry35Ab2 (Accession # AAK64563); Cry35Ab3(Accession # AY536891); Cry35Ac1 (Accession # AAG50117); Cry35Ba1(Accession # AAK64566); Cry35Ba2 (Accession # AAT29027); Cry35Ba3(Accession # AAT29026); Cry36Aa1 (Accession # AAK64558); Cry37Aa1(Accession # AAF76376); Cry38Aa1 (Accession # AAK64559); Cry39Aa1(Accession # BAB72016); Cry40Aa1 (Accession # BAB72018); Cry40Ba1(Accession # BAC77648); Cry40Ca1 (Accession # EU381045); Cry40Da1(Accession # ACF15199); Cry41Aa1 (Accession # BAD35157); Cry41Ab1(Accession # BAD35163); Cry41Ba1 (Accession # HM461871); Cry41Ba2(Accession # ZP_04099652); Cry42Aa1 (Accession # BAD35166); Cry43Aa1(Accession # BAD15301); Cry43Aa2 (Accession # BAD95474); Cry43Ba1(Accession # BAD15303); Cry43Ca1 (Accession # KC156676); Cry43Cb1(Accession # KC156695); Cry43Cc1 (Accession # KC156696); Cry43-like(Accession # BAD15305); Cry44Aa (Accession # BAD08532); Cry45Aa(Accession # BAD22577); Cry46Aa (Accession # BAC79010); Cry46Aa2(Accession # BAG68906); Cry46Ab (Accession # BAD35170); Cry47Aa(Accession # AAY24695); Cry48Aa (Accession # CAJ18351); Cry48Aa2(Accession # CAJ86545); Cry48Aa3 (Accession # CAJ86546); Cry48Ab(Accession # CAJ86548); Cry48Ab2 (Accession # CAJ86549); Cry49Aa(Accession # CAH56541); Cry49Aa2 (Accession # CAJ86541); Cry49Aa3(Accession # CAJ86543); Cry49Aa4 (Accession # CAJ86544); Cry49Ab1(Accession # CAJ86542); Cry50Aa1 (Accession # BAE86999); Cry50Ba1(Accession # GU446675); Cry50Ba2 (Accession # GU446676); Cry51Aa1(Accession # ABI14444); Cry51Aa2 (Accession # GU570697); Cry52Aa1(Accession # EF613489); Cry52Ba1 (Accession # FJ361760); Cry53Aa1(Accession # EF633476); Cry53Ab1 (Accession # FJ361759); Cry54Aa1(Accession # ACA52194); Cry54Aa2 (Accession # GQ140349); Cry54Ba1(Accession # GU446677); Cry55Aa1 (Accession # ABW88932); Cry54Ab1(Accession # JQ916908); Cry55Aa2 (Accession # AAE33526); Cry56Aa1(Accession # ACU57499); Cry56Aa2 (Accession # GQ483512); Cry56Aa3(Accession # JX025567); Cry57Aa1 (Accession # ANC87261); Cry58Aa1(Accession # ANC87260); Cry59Ba1 (Accession # JN790647); Cry59Aa1(Accession # ACR43758); Cry60Aa1 (Accession # ACU24782); Cry60Aa2(Accession # EAO57254); Cry60Aa3 (Accession # EEM99278); Cry60Ba1(Accession # GU810818); Cry60Ba2 (Accession # EAO57253); Cry60Ba3(Accession # EEM99279); Cry61Aa1 (Accession # HM035087); Cry61Aa2(Accession # HM132125); Cry61Aa3 (Accession # EEM19308); Cry62Aa1(Accession # HM054509); Cry63Aa1 (Accession # BA144028); Cry64Aa1(Accession # BAJ05397); Cry65Aa1 (Accession # HM461868); Cry65Aa2(Accession # ZP_04123838); Cry66Aa1 (Accession # HM485581); Cry66Aa2(Accession # ZP_04099945); Cry67Aa1 (Accession # HM485582); Cry67Aa2(Accession # ZP_04148882); Cry68Aa1 (Accession # HQ113114); Cry69Aa1(Accession # HQ401006); Cry69Aa2 (Accession # JQ821388); Cry69Ab1(Accession # JN209957); Cry70Aa1 (Accession # JN646781); Cry70Ba1(Accession # ADO51070); Cry70Bb1 (Accession # EEL67276); Cry71Aa1(Accession # JX025568); Cry72Aa1 (Accession # JX025569).

Examples of 6-endotoxins also include but are not limited to Cry1Aproteins of U.S. Pat. Nos. 5,880,275 and 7,858,849; a DIG-3 or DIG-11toxin (N-terminal deletion of α-helix 1 and/or α-helix 2 variants of Cryproteins such as Cry1A) of U.S. Pat. Nos. 8,304,604 and 8,304,605, Cry1Bof U.S. patent application Ser. No. 10/525,318; Cry1C of U.S. Pat. No.6,033,874; Cry1F of U.S. Pat. Nos. 5,188,960, 6,218,188; Cry1A/Fchimeras of U.S. Pat. Nos. 7,070,982; 6,962,705 and 6,713,063); a Cry2protein such as Cry2Ab protein of U.S. Pat. No. 7,064,249); a Cry3Aprotein including but not limited to an engineered hybrid insecticidalprotein (eHIP) created by fusing unique combinations of variable regionsand conserved blocks of at least two different Cry proteins (US PatentApplication Publication Number 2010/0017914); a Cry4 protein; a Cry5protein; a Cry6 protein; Cry8 proteins of U.S. Pat. Nos. 7,329,736,7,449,552, 7,803,943, 7,476,781, 7,105,332, 7,378,499 and 7,462,760; aCry9 protein such as such as members of the Cry9A, Cry9B, Cry9C, Cry9D,Cry9E, and Cry9F families; a Cry15 protein of Naimov, et al., (2008)Applied and Environmental Microbiology 74:7145-7151; a Cry22, a Cry34Ab1protein of U.S. Pat. Nos. 6,127,180, 6,624,145 and 6,340,593; a CryET33and CryET34 protein of U.S. Pat. Nos. 6,248,535, 6,326,351, 6,399,330,6,949,626, 7,385,107 and 7,504,229; a CryET33 and CryET34 homologs of USPatent Publication Numbers 2006/0191034, 2012/0278954, and PCTPublication Number WO 2012/139004; a Cry35Ab1 protein of U.S. Pat. Nos.6,083,499, 6,548,291 and 6,340,593; a Cry46 protein, a Cry 51 protein, aCry binary toxin; a TIC901 or related toxin; TIC807 of US 2008/0295207;ET29, ET37, TIC809, TIC810, TIC812, TIC127, TIC128 of PCT US2006/033867; AXMI-027, AXMI-036, and AXMI-038 of U.S. Pat. No.8,236,757; AXMI-031, AXMI-039, AXMI-040, AXMI-049 of U.S. Pat. No.7,923,602; AXMI-018, AXMI-020, and AXMI-021 of WO 2006/083891; AXMI-010of WO 2005/038032; AXMI-003 of WO 2005/021585; AXMI-008 of US2004/0250311; AXMI-006 of US 2004/0216186; AXMI-007 of US 2004/0210965;AXMI-009 of US 2004/0210964; AXMI-014 of US 2004/0197917; AXMI-004 of US2004/0197916; AXMI-028 and AXMI-029 of WO 2006/119457; AXMI-007,AXMI-008, AXMI-0080rf2, AXMI-009, AXMI-014 and AXMI-004 of WO2004/074462; AXMI-150 of U.S. Pat. No. 8,084,416; AXMI-205 ofUS20110023184; AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-019,AXMI-044, AXMI-037, AXMI-043, AXMI-033, AXMI-034, AXMI-022, AXMI-023,AXMI-041, AXMI-063, and AXMI-064 of US 2011/0263488; AXMI-R1 and relatedproteins of US 2010/0197592; AXMI221Z, AXMI222z, AXMI223z, AXMI224z andAXMI225z of WO 2011/103248; AXMI218, AXMI219, AXMI220, AXMI226, AXMI227,AXMI228, AXMI229, AXMI230, and AXMI231 of WO11/103247; AXMI-115,AXMI-113, AXMI-005, AXMI-163 and AXMI-184 of U.S. Pat. No. 8,334,431;AXMI-001, AXMI-002, AXMI-030, AXMI-035, and AXMI-045 of US 2010/0298211;AXMI-066 and AXMI-076 of US20090144852; AXMI128, AXMI130, AXMI131,AXMI133, AXMI140, AXMI141, AXMI142, AXMI143, AXMI144, AXMI146, AXMI148,AXMI149, AXMI152, AXMI153, AXMI154, AXMI155, AXMI156, AXMI157, AXMI158,AXMI162, AXMI165, AXMI166, AXMI167, AXMI168, AXMI169, AXMI170, AXMI171,AXMI172, AXMI173, AXMI174, AXMI175, AXMI176, AXMI177, AXMI178, AXMI179,AXMI180, AXMI181, AXMI182, AXMI185, AXMI186, AXMI187, AXMI188, AXMI189of U.S. Pat. No. 8,318,900; AXMI079, AXMI080, AXMI081, AXMI082, AXMI091,AXMI092, AXMI096, AXMI097, AXMI098, AXMI099, AXMI100, AXMI101, AXMI102,AXMI103, AXMI104, AXMI107, AXMI108, AXMI109, AXMI110, AXMI111, AXMI112,AXMI114, AXMI116, AXMI117, AXMI118, AXMI119, AXMI120, AXMI121, AXMI122,AXMI123, AXMI124, AXMI1257, AXMI1268, AXMI127, AXMI129, AXMI164,AXMI151, AXMI161, AXMI183, AXMI132, AXMI138, AXMI137 of US 2010/0005543;and Cry proteins such as Cry1A and Cry3A having modified proteolyticsites of U.S. Pat. No. 8,319,019; and a Cry1Ac, Cry2Aa and Cry1Ca toxinprotein from Bacillus thuringiensis strain VBTS 2528 of US PatentApplication Publication Number 2011/0064710. Other Cry proteins are wellknown to one skilled in the art (see, Crickmore, et al., “Bacillusthuringiensis toxin nomenclature” (2011), atlifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/which can be accessed on theworld-wide web using the “www” prefix). The insecticidal activity of Cryproteins is well known to one skilled in the art (for review, see, vanFrannkenhuyzen, (2009) J. Invert. Path. 101:1-16). The use of Cryproteins as transgenic plant traits is well known to one skilled in theart and Cry-transgenic plants including but not limited to Cry1Ac,Cry1Ac+Cry2Ab, Cry1Ab, Cry1A.105, Cry1F, Cry1Fa2, Cry1F+Cry1Ac, Cry2Ab,Cry3A, mCry3A, Cry3Bb1, Cry34Ab1, Cry35Ab1, Vip3A, mCry3A, Cry9c andCBI-Bt have received regulatory approval (see, Sanahuja, (2011) PlantBiotech Journal 9:283-300 and the CERA (2010) GM Crop Database Centerfor Environmental Risk Assessment (CERA), ILSI Research Foundation,Washington D.C. at cera-gmc.org/index.php?action=gm_crop_database whichcan be accessed on the world-wide web using the “www” prefix). More thanone pesticidal proteins well known to one skilled in the art can also beexpressed in plants such as Vip3Ab & Cry1Fa (US2012/0317682), Cry1BE &Cry1F (US2012/0311746), Cry1CA & Cry1AB (US2012/0311745), Cry1F & CryCa(US2012/0317681), Cry1DA & Cry1BE (US2012/0331590), Cry1DA & Cry1Fa(US2012/0331589), Cry1AB & Cry1BE (US2012/0324606), and Cry1Fa & Cry2Aa,Cry1I or Cry1E (US2012/0324605). Pesticidal proteins also includeinsecticidal lipases including lipid acyl hydrolases of U.S. Pat. No.7,491,869, and cholesterol oxidases such as from Streptomyces (Purcellet al. (1993) Biochem Biophys Res Commun 15:1406-1413). Pesticidalproteins also include VIP (vegetative insecticidal proteins) toxins ofU.S. Pat. Nos. 5,877,012, 6,107,279, 6,137,033, 7,244,820, 7,615,686,and 8,237,020, and the like. Other VIP proteins are well known to oneskilled in the art (see,lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/vip.html which can beaccessed on the world-wide web using the “www” prefix). Pesticidalproteins also include toxin complex (TC) proteins, obtainable fromorganisms such as Xenorhabdus, Photorhabdus and Paenibacillus (see, U.S.Pat. Nos. 7,491,698 and 8,084,418). Some TC proteins have “stand alone”insecticidal activity and other TC proteins enhance the activity of thestand-alone toxins produced by the same given organism. The toxicity ofa “stand-alone” TC protein (from Photorhabdus, Xenorhabdus orPaenibacillus, for example) can be enhanced by one or more TC protein“potentiators” derived from a source organism of a different genus.There are three main types of TC proteins. As referred to herein, ClassA proteins (“Protein A”) are stand-alone toxins. Class B proteins(“Protein B”) and Class C proteins (“Protein C”) enhance the toxicity ofClass A proteins. Examples of Class A proteins are TcbA, TcdA, XptA1 andXptA2. Examples of Class B proteins are TcaC, TcdB, XptB1Xb and XptC1Wi.Examples of Class C proteins are TccC, XptC1Xb and XptB1Wi. Pesticidalproteins also include spider, snake and scorpion venom proteins.Examples of spider venom peptides include but are not limited tolycotoxin-1 peptides and mutants thereof (U.S. Pat. No. 8,334,366).

(C) A polynucleotide encoding an insect-specific hormone or pheromonesuch as an ecdysteroid and juvenile hormone, a variant thereof, amimetic based thereon or an antagonist or agonist thereof. See, forexample, the disclosure by Hammock, et al., (1990) Nature 344:458, ofbaculovirus expression of cloned juvenile hormone esterase, aninactivator of juvenile hormone.

(D) A polynucleotide encoding an insect-specific peptide which, uponexpression, disrupts the physiology of the affected pest. For example,see the disclosures of, Regan, (1994) J. Biol. Chem. 269:9 (expressioncloning yields DNA coding for insect diuretic hormone receptor); Pratt,et al., (1989) Biochem. Biophys. Res. Comm. 163:1243 (an allostatin isidentified in Diploptera puntata); Chattopadhyay, et al., (2004)Critical Reviews in Microbiology 30(1):33-54; Zjawiony, (2004) J NatProd 67(2):300-310; Carlini and Grossi-de-Sa, (2002) Toxicon40(11):1515-1539; Ussuf, et al., (2001) Curr Sci. 80(7):847-853 andVasconcelos and Oliveira, (2004) Toxicon 44(4):385-403. See also, U.S.Pat. No. 5,266,317 to Tomalski, et al., who disclose genes encodinginsect-specific toxins.

(E) A polynucleotide encoding an enzyme responsible for ahyperaccumulation of a monoterpene, a sesquiterpene, a steroid,hydroxamic acid, a phenylpropanoid derivative or another non-proteinmolecule with insecticidal activity.

(F) A polynucleotide encoding an enzyme involved in the modification,including the post-translational modification, of a biologically activemolecule; for example, a glycolytic enzyme, a proteolytic enzyme, alipolytic enzyme, a nuclease, a cyclase, a transaminase, an esterase, ahydrolase, a phosphatase, a kinase, a phosphorylase, a polymerase, anelastase, a chitinase and a glucanase, whether natural or synthetic.See, PCT Application WO 1993/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase gene. DNA molecules whichcontain chitinase-encoding sequences can be obtained, for example, fromthe ATCC® under Accession Numbers 39637 and 67152. See also, Kramer, etal., (1993) Insect Biochem. Molec. Biol. 23:691, who teach thenucleotide sequence of a cDNA encoding tobacco hookworm chitinase andKawalleck, et al., (1993) Plant Molec. Biol. 21:673, who provide thenucleotide sequence of the parsley ubi4-2 polyubiquitin gene, and U.S.Pat. Nos. 6,563,020; 7,145,060 and 7,087,810.

(G) A polynucleotide encoding a molecule that stimulates signaltransduction. For example, see the disclosure by Botella, et al., (1994)Plant Molec. Biol. 24:757, of nucleotide sequences for mung beancalmodulin cDNA clones, and Griess, et al., (1994) Plant Physiol.104:1467, who provide the nucleotide sequence of a maize calmodulin cDNAclone.

(H) A polynucleotide encoding a hydrophobic moment peptide. See, PCTApplication WO 1995/16776 and U.S. Pat. No. 5,580,852 disclosure ofpeptide derivatives of Tachyplesin which inhibit fungal plant pathogens)and PCT Application WO 1995/18855 and U.S. Pat. No. 5,607,914 (teachessynthetic antimicrobial peptides that confer disease resistance).

(I) A polynucleotide encoding a membrane permease, a channel former or achannel blocker. For example, see the disclosure by Jaynes, et al.,(1993) Plant Sci. 89:43, of heterologous expression of a cecropin-betalytic peptide analog to render transgenic tobacco plants resistant toPseudomonas solanacearum.

(J) A gene encoding a viral-invasive protein or a complex toxin derivedtherefrom. For example, the accumulation of viral coat proteins intransformed plant cells imparts resistance to viral infection and/ordisease development effected by the virus from which the coat proteingene is derived, as well as by related viruses. See, Beachy, et al.,(1990) Ann. Rev. Phytopathol. 28:451. Coat protein-mediated resistancehas been conferred upon transformed plants against alfalfa mosaic virus,cucumber mosaic virus, tobacco streak virus, potato virus X, potatovirus Y, tobacco etch virus, tobacco rattle virus and tobacco mosaicvirus. Id.

(K) A gene encoding an insect-specific antibody or an immunotoxinderived therefrom. Thus, an antibody targeted to a critical metabolicfunction in the insect gut would inactivate an affected enzyme, killingthe insect. Cf. Taylor, et al., Abstract #497, SEVENTH INT'L SYMPOSIUMON MOLECULAR PLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994)(enzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments).

(L) A gene encoding a virus-specific antibody. See, for example,Tavladoraki, et al., (1993) Nature 366:469, who show that transgenicplants expressing recombinant antibody genes are protected from virusattack.

(M) A polynucleotide encoding a developmental-arrestive protein producedin nature by a pathogen or a parasite. Thus, fungal endoalpha-1,4-D-polygalacturonases facilitate fungal colonization and plantnutrient release by solubilizing plant cell wallhomo-alpha-1,4-D-galacturonase. See, Lamb, et al., (1992) Bio/Technology10:1436. The cloning and characterization of a gene which encodes a beanendopolygalacturonase-inhibiting protein is described by Toubart, etal., (1992) Plant J. 2:367.

(N) A polynucleotide encoding a developmental-arrestive protein producedin nature by a plant. For example, Logemann, et al., (1992)Bio/Technology 10:305, have shown that transgenic plants expressing thebarley ribosome-inactivating gene have an increased resistance to fungaldisease.

(O) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, (1995) Current Biology5(2), Pieterse and Van Loon, (2004) Curr. Opin. Plant Bio. 7(4):456-64and Somssich, (2003) Cell 113(7):815-6.

(P) Antifungal genes (Cornelissen and Melchers, (1993) Pl. Physiol.101:709-712 and Parijs, et al., (1991) Planta 183:258-264 and Bushnell,et al., (1998) Can. J. of Plant Path. 20(2):137-149. Also see, U.S.patent application Ser. Nos. 09/950,933; 11/619,645; 11/657,710;11/748,994; 11/774,121 and U.S. Pat. Nos. 6,891,085 and 7,306,946. LysMReceptor-like kinases for the perception of chitin fragments as a firststep in plant defense response against fungal pathogens (US2012/0110696).

(Q) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see, U.S. Pat. Nos. 5,716,820; 5,792,931; 5,798,255;5,846,812; 6,083,736; 6,538,177; 6,388,171 and 6,812,380.

(R) A polynucleotide encoding a Cystatin and cysteine proteinaseinhibitors. See, U.S. Pat. No. 7,205,453.

(S) Defensin genes. See, WO 2003/000863 and U.S. Pat. Nos. 6,911,577;6,855,865; 6,777,592 and 7,238,781.

(T) Genes conferring resistance to nematodes. See, e.g., PCT ApplicationWO 1996/30517; PCT Application WO 1993/19181, WO 2003/033651 and Urwin,et al., (1998) Planta 204:472-479, Williamson, (1999) Curr Opin PlantBio. 2(4):327-31; U.S. Pat. Nos. 6,284,948 and 7,301,069 and miR164genes (WO 2012/058266).

(U) Genes that confer resistance to Phytophthora Root Rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker, et al., Phytophthora Root Rot ResistanceGene Mapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

(V) Genes that confer resistance to Brown Stem Rot, such as described inU.S. Pat. No. 5,689,035 and incorporated by reference for this purpose.

(W) Genes that confer resistance to Colletotrichum, such as described inUS Patent Application Publication US 2009/0035765 and incorporated byreference for this purpose. This includes the Rcg locus that may beutilized as a single locus conversion.

2. Transgenes that Confer Resistance to a Herbicide, for Example:

(A) A polynucleotide encoding resistance to a herbicide that inhibitsthe growing point or meristem, such as an imidazolinone or asulfonylurea. Exemplary genes in this category code for mutant ALS andAHAS enzyme as described, for example, by Lee, et al., (1988) EMBO J.7:1241 and Miki, et al., (1990) Theor. Appl. Genet. 80:449,respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870;5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107; 5,928,937 and5,378,824; U.S. patent application Ser. No. 11/683,737 and InternationalPublication WO 1996/33270.

(B) A polynucleotide encoding a protein for resistance to Glyphosate(resistance imparted by mutant 5-enolpyruvl-3-phosphikimate synthase(EPSP) and aroA genes, respectively) and other phosphono compounds suchas glufosinate (phosphinothricin acetyl transferase (PAT) andStreptomyces hygroscopicus phosphinothricin acetyl transferase (bar)genes), and pyridinoxy or phenoxy proprionic acids and cyclohexones(ACCase inhibitor-encoding genes). See, for example, U.S. Pat. No.4,940,835 to Shah, et al., which discloses the nucleotide sequence of aform of EPSPS which can confer glyphosate resistance. U.S. Pat. No.5,627,061 to Barry, et al., also describes genes encoding EPSPS enzymes.See also, U.S. Pat. Nos. 6,566,587; 6,338,961; 6,248,876 B1; 6,040,497;5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642;5,094,945, 4,940,835; 5,866,775; 6,225,114 B1; 6,130,366; 5,310,667;4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE 37,287 E and5,491,288 and International Publications EP 1173580; WO 2001/66704; EP1173581 and EP 1173582, which are incorporated herein by reference forthis purpose. Glyphosate resistance is also imparted to plants thatexpress a gene encoding a glyphosate oxido-reductase enzyme as describedmore fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, which areincorporated herein by reference for this purpose. In additionglyphosate resistance can be imparted to plants by the over expressionof genes encoding glyphosate N-acetyltransferase. See, for example, U.S.Pat. Nos. 7,462,481; 7,405,074 and US Patent Application PublicationNumber US 2008/0234130. A DNA molecule encoding a mutant aroA gene canbe obtained under ATCC® Accession Number 39256, and the nucleotidesequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061 toComai. EP Application Number 0 333 033 to Kumada, et al., and U.S. Pat.No. 4,975,374 to Goodman, et al., disclose nucleotide sequences ofglutamine synthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EP ApplicationNumbers 0 242 246 and 0 242 236 to Leemans, et al.; De Greef, et al.,(1989) Bio/Technology 7:61, describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. See also, U.S. Pat. Nos. 5,969,213; 5,489,520;5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024;6,177,616 B1 and 5,879,903, which are incorporated herein by referencefor this purpose. Exemplary genes conferring resistance to phenoxyproprionic acids and cyclohexones, such as sethoxydim and haloxyfop, arethe Acc1-S1, Acc1-52 and Acc1-53 genes described by Marshall, et al.,(1992) Theor. Appl. Genet. 83:435.

(C) A polynucleotide encoding a protein for resistance to herbicide thatinhibits photosynthesis, such as a triazine (psbA and gs+ genes) and abenzonitrile (nitrilase gene). Przibilla, et al., (1991) Plant Cell3:169, describe the transformation of Chlamydomonas with plasmidsencoding mutant psbA genes. Nucleotide sequences for nitrilase genes aredisclosed in U.S. Pat. No. 4,810,648 to Stalker and DNA moleculescontaining these genes are available under ATCC® Accession Numbers53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes, et al., (1992) Biochem.J. 285:173.

(D) A polynucleotide encoding a protein for resistance to Acetohydroxyacid synthase, which has been found to make plants that express thisenzyme resistant to multiple types of herbicides, has been introducedinto a variety of plants (see, e.g., Hattori, et al., (1995) Mol GenGenet. 246:419). Other genes that confer resistance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota, et al., (1994)Plant Physiol 106:17), genes for glutathione reductase and superoxidedismutase (Aono, et al., (1995) Plant Cell Physiol 36:1687) and genesfor various phosphotransferases (Datta, et al., (1992) Plant Mol Biol20:619).

(E) A polynucleotide encoding resistance to a herbicide targetingProtoporphyrinogen oxidase (protox) which is necessary for theproduction of chlorophyll. The protox enzyme serves as the target for avariety of herbicidal compounds. These herbicides also inhibit growth ofall the different species of plants present, causing their totaldestruction. The development of plants containing altered protoxactivity which are resistant to these herbicides are described in U.S.Pat. Nos. 6,288,306 B1; 6,282,837 B1 and 5,767,373 and InternationalPublication WO 2001/12825.

(F) The aad-1 gene (originally from Sphingobium herbicidovorans) encodesthe aryloxyalkanoate dioxygenase (AAD-1) protein. The trait conferstolerance to 2,4-dichlorophenoxyacetic acid and aryloxyphenoxypropionate(commonly referred to as “fop” herbicides such as quizalofop)herbicides. The aad-1 gene, itself, for herbicide tolerance in plantswas first disclosed in WO 2005/107437 (see also, US 2009/0093366). Theaad-12 gene, derived from Delftia acidovorans, which encodes thearyloxyalkanoate dioxygenase (AAD-12) protein that confers tolerance to2,4-dichlorophenoxyacetic acid and pyridyloxyacetate herbicides bydeactivating several herbicides with an aryloxyalkanoate moiety,including phenoxy auxin (e.g., 2,4-D, MCPA), as well as pyridyloxyauxins (e.g., fluroxypyr, triclopyr).

(G) A polynucleotide encoding a herbicide resistant dicambamonooxygenase disclosed in US Patent Application Publication2003/0135879 for imparting dicamba tolerance;

(H) A polynucleotide molecule encoding bromoxynil nitrilase (Bxn)disclosed in U.S. Pat. No. 4,810,648 for imparting bromoxynil tolerance;

(I) A polynucleotide molecule encoding phytoene (crtl) described inMisawa, et al., (1993) Plant J. 4:833-840 and in Misawa, et al., (1994)Plant J. 6:481-489 for norflurazon tolerance.

3. Transgenes that Confer or Contribute to an Altered GrainCharacteristic

Such as:

(A) Altered fatty acids, for example, by

(1) Down-regulation of stearoyl-ACP to increase stearic acid content ofthe plant. See, Knultzon, et al., (1992) Proc. Natl. Acad. Sci. USA89:2624 and WO 1999/64579 (Genes to Alter Lipid Profiles in Corn).

(2) Elevating oleic acid via FAD-2 gene modification and/or decreasinglinolenic acid via FAD-3 gene modification (see, U.S. Pat. Nos.6,063,947; 6,323,392; 6,372,965 and WO 1993/11245).

(3) Altering conjugated linolenic or linoleic acid content, such as inWO 2001/12800.

(4) Altering LEC1, AGP, Dek1, Superal1, mi1 ps, various Ipa genes suchas Ipa1, Ipa3, hpt or hggt. For example, see, WO 2002/42424, WO1998/22604, WO 2003/011015, WO 2002/057439, WO 2003/011015, U.S. Pat.Nos. 6,423,886, 6,197,561, 6,825,397 and US Patent ApplicationPublication Numbers US 2003/0079247, US 2003/0204870 and Rivera-Madrid,et al., (1995) Proc. Natl. Acad. Sci. 92:5620-5624.

(5) Genes encoding delta-8 desaturase for making long-chainpolyunsaturated fatty acids (U.S. Pat. Nos. 8,058,571 and 8,338,152),delta-9 desaturase for lowering saturated fats (U.S. Pat. No.8,063,269), Primula Δ6-desaturase for improving omega-3 fatty acidprofiles.

(6) Isolated nucleic acids and proteins associated with lipid and sugarmetabolism regulation, in particular, lipid metabolism protein (LMP)used in methods of producing transgenic plants and modulating levels ofseed storage compounds including lipids, fatty acids, starches or seedstorage proteins and use in methods of modulating the seed size, seednumber, seed weights, root length and leaf size of plants (EP 2404499).

(7) Altering expression of a High-Level Expression of Sugar-Inducible 2(HSI2) protein in the plant to increase or decrease expression of HSI2in the plant. Increasing expression of HSI2 increases oil content whiledecreasing expression of HSI2 decreases abscisic acid sensitivity and/orincreases drought resistance (US Patent Application Publication Number2012/0066794).

(8) Expression of cytochrome b5 (Cb5) alone or with FAD2 to modulate oilcontent in plant seed, particularly to increase the levels of omega-3fatty acids and improve the ratio of omega-6 to omega-3 fatty acids (USPatent Application Publication Number 2011/0191904).

(9) Nucleic acid molecules encoding wrinkled1-like polypeptides formodulating sugar metabolism (U.S. Pat. No. 8,217,223).

(B) Altered phosphorus content, for example, by the

(1) Introduction of a phytase-encoding gene would enhance breakdown ofphytate, adding more free phosphate to the transformed plant. Forexample, see, Van Hartingsveldt, et al., (1993) Gene 127:87, for adisclosure of the nucleotide sequence of an Aspergillus niger phytasegene.

(2) Modulating a gene that reduces phytate content. In maize, this, forexample, could be accomplished, by cloning and then re-introducing DNAassociated with one or more of the alleles, such as the LPA alleles,identified in maize mutants characterized by low levels of phytic acid,such as in WO 2005/113778 and/or by altering inositol kinase activity asin WO 2002/059324, US Patent Application Publication Number2003/0009011, WO 2003/027243, US Patent Application Publication Number2003/0079247, WO 1999/05298, U.S. Pat. Nos. 6,197,561, 6,291,224,6,391,348, WO 2002/059324, US Patent Application Publication Number2003/0079247, WO 1998/45448, WO 1999/55882, WO 2001/04147.

(C) Altered carbohydrates affected, for example, by altering a gene foran enzyme that affects the branching pattern of starch or, a genealtering thioredoxin such as NTR and/or TRX (see, U.S. Pat. No.6,531,648. which is incorporated by reference for this purpose) and/or agamma zein knock out or mutant such as cs27 or TUSC27 or en27 (see, U.S.Pat. No. 6,858,778 and US Patent Application Publication Number2005/0160488, US Patent Application Publication Number 2005/0204418,which are incorporated by reference for this purpose). See, Shiroza, etal., (1988) J. Bacteriol. 170:810 (nucleotide sequence of Streptococcusmutant fructosyltransferase gene), Steinmetz, et al., (1985) Mol. Gen.Genet. 200:220 (nucleotide sequence of Bacillus subtilis levansucrasegene), Pen, et al., (1992) Bio/Technology 10:292 (production oftransgenic plants that express Bacillus licheniformis alpha-amylase),Elliot, et al., (1993) Plant Molec. Biol. 21:515 (nucleotide sequencesof tomato invertase genes), Søgaard, et al., (1993) J. Biol. Chem.268:22480 (site-directed mutagenesis of barley alpha-amylase gene) andFisher, et al., (1993) Plant Physiol. 102:1045 (maize endosperm starchbranching enzyme II), WO 1999/10498 (improved digestibility and/orstarch extraction through modification of UDP-D-xylose 4-epimerase,Fragile 1 and 2, Ref1, HCHL, C4H), U.S. Pat. No. 6,232,529 (method ofproducing high oil seed by modification of starch levels (AGP)). Thefatty acid modification genes mentioned herein may also be used toaffect starch content and/or composition through the interrelationshipof the starch and oil pathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683,US Patent Application Publication Number 2004/0034886 and WO 2000/68393involving the manipulation of antioxidant levels and WO 2003/082899through alteration of a homogentisate geranyl geranyl transferase(hggt).

(E) Altered essential seed amino acids. For example, see, U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO 1999/40209 (alteration of amino acid compositions inseeds), WO 1999/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO 1998/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO 1998/56935 (plant amino acid biosyntheticenzymes), WO 1998/45458 (engineered seed protein having higherpercentage of essential amino acids), WO 1998/42831 (increased lysine),U.S. Pat. No. 5,633,436 (increasing sulfur amino acid content), U.S.Pat. No. 5,559,223 (synthetic storage proteins with defined structurecontaining programmable levels of essential amino acids for improvementof the nutritional value of plants), WO 1996/01905 (increasedthreonine), WO 1995/15392 (increased lysine), US Patent ApplicationPublication Number 2003/0163838, US Patent Application PublicationNumber 2003/0150014, US Patent Application Publication Number2004/0068767, U.S. Pat. No. 6,803,498, WO 2001/79516.

4. Genes that Control Male-Sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar, et al., and chromosomal translocationsas described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen, et al., U.S. Pat. No. 5,432,068,describe a system of nuclear male sterility which includes: identifyinga gene which is critical to male fertility; silencing this native genewhich is critical to male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 2001/29237).

(B) Introduction of various stamen-specific promoters (WO 1992/13956, WO1992/13957).

(C) Introduction of the barnase and the barstar gene (Paul, et al.,(1992) Plant Mol. Biol. 19:611-622).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014 and 6,265,640, all of which are hereby incorporatedby reference.

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see, Lyznik, et al., (2003) Plant Cell Rep 21:925-932 andWO 1999/25821, which are hereby incorporated by reference. Other systemsthat may be used include the Gin recombinase of phage Mu (Maeser, etal., (1991) Vicki Chandler, The Maize Handbook ch. 118 (Springer-Verlag1994), the Pin recombinase of E. coli (Enomoto, et al., 1983) and theR/RS system of the pSRi plasmid (Araki, et al., 1992).

6. Genes that Affect Abiotic Stress Resistance

Including but not limited to flowering, ear and seed development,enhancement of nitrogen utilization efficiency, altered nitrogenresponsiveness, drought resistance or tolerance, cold resistance ortolerance and salt resistance or tolerance and increased yield understress.

(A) For example, see: WO 2000/73475 where water use efficiency isaltered through alteration of malate; U.S. Pat. Nos. 5,892,009,5,965,705, 5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866,6,717,034, 6,801,104, WO 2000/060089, WO 2001/026459, WO 2001/035725, WO2001/034726, WO 2001/035727, WO 2001/036444, WO 2001/036597, WO2001/036598, WO 2002/015675, WO 2002/017430, WO 2002/077185, WO2002/079403, WO 2003/013227, WO 2003/013228, WO 2003/014327, WO2004/031349, WO 2004/076638, WO 199809521.

(B) WO 199938977 describing genes, including CBF genes and transcriptionfactors effective in mitigating the negative effects of freezing, highsalinity and drought on plants, as well as conferring other positiveeffects on plant phenotype.

(C) US Patent Application Publication Number 2004/0148654 and WO2001/36596 where abscisic acid is altered in plants resulting inimproved plant phenotype such as increased yield and/or increasedtolerance to abiotic stress.

(D) WO 2000/006341, WO 2004/090143, U.S. Pat. Nos. 7,531,723 and6,992,237 where cytokinin expression is modified resulting in plantswith increased stress tolerance, such as drought tolerance, and/orincreased yield. Also see, WO 2002/02776, WO 2003/052063, JP2002/281975, U.S. Pat. No. 6,084,153, WO 2001/64898, U.S. Pat. Nos.6,177,275 and 6,107,547 (enhancement of nitrogen utilization and alterednitrogen responsiveness).

(E) For ethylene alteration, see, US Patent Application PublicationNumber 2004/0128719, US Patent Application Publication Number2003/0166197 and WO 2000/32761.

(F) For plant transcription factors or transcriptional regulators ofabiotic stress, see, e.g., US Patent Application Publication Number2004/0098764 or US Patent Application Publication Number 2004/0078852.

(G) Genes that increase expression of vacuolar pyrophosphatase such asAVP1 (U.S. Pat. No. 8,058,515) for increased yield; nucleic acidencoding a HSFA4 or a HSFA5 (Heat Shock Factor of the class A4 or A5)polypeptides, an oligopeptide transporter protein (OPT4-like)polypeptide; a plastochron2-like (PLA2-like) polypeptide or a Wuschelrelated homeobox 1-like (WOX1-like) polypeptide (U. Patent ApplicationPublication Number US 2011/0283420).

(H) Down regulation of polynucleotides encoding poly (ADP-ribose)polymerase (PARP) proteins to modulate programmed cell death (U.S. Pat.No. 8,058,510) for increased vigor.

(I) Polynucleotide encoding DTP21 polypeptides for conferring droughtresistance (US Patent Application Publication Number US 2011/0277181).

(J) Nucleotide sequences encoding ACC Synthase 3 (ACS3) proteins formodulating development, modulating response to stress, and modulatingstress tolerance (US Patent Application Publication Number US2010/0287669).

(K) Polynucleotides that encode proteins that confer a drought tolerancephenotype (DTP) for conferring drought resistance (WO 2012/058528).

(L) Tocopherol cyclase (TC) genes for conferring drought and salttolerance (US Patent Application Publication Number 2012/0272352).

(M) CAAX amino terminal family proteins for stress tolerance (U.S. Pat.No. 8,338,661).

(N) Mutations in the SAL1 encoding gene have increased stress tolerance,including increased drought resistant (US Patent Application PublicationNumber 2010/0257633).

(O) Expression of a nucleic acid sequence encoding a polypeptideselected from the group consisting of: GRF polypeptide, RAA1-likepolypeptide, SYR polypeptide, ARKL polypeptide, and YTP polypeptideincreasing yield-related traits (US Patent Application PublicationNumber 2011/0061133).

(P) Modulating expression in a plant of a nucleic acid encoding a ClassIII Trehalose Phosphate Phosphatase (TPP) polypeptide for enhancingyield-related traits in plants, particularly increasing seed yield (USPatent Application Publication Number 2010/0024067).

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see e.g., WO1997/49811 (LHY), WO 1998/56918 (ESD4), WO 1997/10339 and U.S. Pat. No.6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT), WO 1996/14414 (CON), WO1996/38560, WO 2001/21822 (VRN1), WO 2000/44918 (VRN2), WO 1999/49064(GI), WO 2000/46358 (FR1), WO 1997/29123, U.S. Pat. Nos. 6,794,560,6,307,126 (GAI), WO 1999/09174 (D8 and Rht) and WO 2004/076638 and WO2004/031349 (transcription factors).

7. Genes that Confer Increased Yield

(A) A transgenic crop plant transformed by a1-AminoCyclopropane-1-Carboxylate Deaminase-like Polypeptide (ACCDP)coding nucleic acid, wherein expression of the nucleic acid sequence inthe crop plant results in the plant's increased root growth, and/orincreased yield, and/or increased tolerance to environmental stress ascompared to a wild type variety of the plant (U.S. Pat. No. 8,097,769).

(B) Over-expression of maize zinc finger protein gene (Zm-ZFP1) using aseed preferred promoter has been shown to enhance plant growth, increasekernel number and total kernel weight per plant (US Patent ApplicationPublication Number 2012/0079623).

(C) Constitutive over-expression of maize lateral organ boundaries (LOB)domain protein (Zm-LOBDP1) has been shown to increase kernel number andtotal kernel weight per plant (US Patent Application Publication Number2012/0079622).

(D) Enhancing yield-related traits in plants by modulating expression ina plant of a nucleic acid encoding a VIM1 (Variant in Methylation1)-like polypeptide or a VTC2-like (GDP-L-galactose phosphorylase)polypeptide or a DUF1685 polypeptide or an ARF6-like (Auxin ResponsiveFactor) polypeptide (WO 2012/038893).

(E) Modulating expression in a plant of a nucleic acid encoding aSte20-like polypeptide or a homologue thereof gives plants havingincreased yield relative to control plants (EP 2431472).

(F) Genes encoding nucleoside diphosphatase kinase (NDK) polypeptidesand homologs thereof for modifying the plant's root architecture (USPatent Application Publication Number 2009/0064373).

8. Genes that Confer Plant Digestibility.

(A) Altering the level of xylan present in the cell wall of a plant bymodulating expression of xylan synthase (U.S. Pat. No. 8,173,866).

In some embodiment the stacked trait may be a trait or event that hasreceived regulatory approval including but not limited to the events inTable 6A-6F.

TABLE 6A Triticum aestivum Wheat Event Company Description AP205CL BASFInc. Selection for a mutagenized version of the enzyme acetohydroxyacidsynthase (AHAS), also known as acetolactate synthase (ALS) oracetolactate pyruvate- lyase. AP602CL BASF Inc. Selection for amutagenized version of the enzyme acetohydroxyacid synthase (AHAS), alsoknown as acetolactate synthase (ALS) or acetolactate pyruvate- lyase.BW255-2, BASF Inc. Selection for a mutagenized version of the BW238-3enzyme acetohydroxyacid synthase (AHAS), also known as acetolactatesynthase (ALS) or acetolactate pyruvate- lyase. BW7 BASF Inc. Toleranceto imidazolinone herbicides induced by chemical mutagenesis of theacetohydroxyacid synthase (AHAS) gene using sodium azide. MON71800Monsanto Glyphosate tolerant wheat variety Company produced by insertinga modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) encodinggene from the soil bacterium Agrobacterium tumefaciens, strain CP4.SWP965001 Cyanamid Selection for a mutagenized version of the Cropenzyme acetohydroxyacid synthase (AHAS), Protection also known asacetolactate synthase (ALS) or acetolactate pyruvate- lyase. Teal 11ABASF Inc. Selection for a mutagenized version of the enzymeacetohydroxyacid synthase (AHAS), also known as acetolactate synthase(ALS) or acetolactate pyruvate- lyase.

TABLE 6B Helianthus annuus Sunflower Event Company Description X81359BASF Inc. Tolerance to imidazolinone herbicides by selection of anaturally occurring mutant.

TABLE 6C Glycine max L. Soybean Event Company Description A2704-12,Bayer Glufosinate ammonium herbicide A2704-21, CropScience tolerantsoybean produced by inserting A5547-35 (Aventis a modifiedphosphinothricin CropScience acetyltransferase (PAT) encoding gene(AgrEvo)) from the soil bacterium Streptomyces viridochromogenes.A5547-127 Bayer Glufosinate ammonium herbicide CropScience tolerantsoybean produced by inserting (Aventis a modified phosphinothricinCropScience acetyltransferase (PAT) encoding gene (AgrEvo)) from thesoil bacterium Streptomyces viridochromogenes. BPS-CV127-9 BASF Inc. Theintroduced csr1-2 gene from Arabidopsis thaliana encodes anacetohydroxyacid synthase protein that confers tolerance toimidazolinone herbicides due to a point mutation that results in asingle amino acid substitution in which the serine residue at position653 is replaced by asparagine (S653N). DP-305423 Pioneer High oleic acidsoybean produced by Hi-Bred inserting additional copies of a Inter-portion of the omega-6 desaturase national encoding gene, gm-fad2-1resulting in Inc. silencing of the endogenous omega-6 desaturase gene(FAD2-1). DP356043 Pioneer Soybean event with two herbicide Hi-Bredtolerance genes: glyphosate Inter- N-acetlytransferase, which detoxifiesnational glyphosate, and a modified Inc. acetolactate synthase (ALS)gene which is tolerant to ALS-inhibiting herbicides. G94-1, DuPont Higholeic acid soybean produced by G94-19, Canada inserting a second copy ofthe fatty G168 Agri- acid desaturase (GmFad2-1) encoding cultural genefrom soybean, which resulted in Products “silencing” of the endogenoushost gene. GTS 40-3-2 Monsanto Glyphosate tolerant soybean varietyCompany produced by inserting a modified5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) encoding gene fromthe soil bacterium Agrobacterium tumefaciens. GU262 Bayer Glufosinateammonium herbicide CropScience tolerant soybean produced by inserting(Aventis a modified phosphinothricin CropScience acetyltransferase (PAT)encoding gene (AgrEvo)) from the soil bacterium Streptomycesviridochromogenes. MON87701 Monsanto Resistance to Lepidopteran pests ofCompany soybean including velvetbean caterpillar (Anticarsia gemmatalis)and soybean looper (Pseudoplusia includens). MON87701 × MonsantoGlyphosate herbicide tolerance through MON89788 Company expression ofthe EPSPS encoding gene from A. tumefaciens strain CP4, and resistanceto Lepidopteran pests of soybean including velvetbean caterpillar(Anticarsia gemmatalis) and soybean looper (Pseudoplusia includens) viaexpression of the Cry1Ac encoding gene from B. thuringiensis. MON89788Monsanto Glyphosate-tolerant soybean produced Company by inserting amodified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) encodingaroA (epsps) gene from Agrobacterium tumefaciens CP4. OT96-15 Agri- Lowlinolenic acid soybean produced culture & through traditionalcross-breeding to Agri-Food incorporate the novel trait from a Canadanaturally occurring fan1 gene mutant that was selected for low linolenicacid. W62, W98 Bayer Glufosinate ammonium herbicide CropScience tolerantsoybean produced by inserting (Aventis a modified phosphinothricinCropScience acetyltransferase (PAT) encoding gene (AgrEvo)) from thesoil bacterium Streptomyces hygroscopicus.

TABLE 6D Medicago sativa Alfalfa Event Company Description J101, J163Monsanto Glyphosate herbicide tolerant alfalfa Company and (lucerne)produced by inserting a Forage gene encoding the enzyme Genetics5-enolypyruvylshikimate-3-phosphate International synthase (EPSPS) fromthe CP4 strain of Agrobacterium tumefaciens.

TABLE 6E Oryza sativa Rice Event Company Description CL121, BASF Inc.Tolerance to the imidazolinone herbicide, CL141, imazethapyr, induced bychemical CFX51 mutagenesis of the acetolactate synthase (ALS) enzymeusing ethyl methanesulfonate (EMS). IMINTA-1, BASF Inc. Tolerance toimidazolinone herbicides IMINTA-4 induced by chemical mutagenesis of theacetolactate synthase (ALS) enzyme using sodium azide. LLRICE06, AventisGlufosinate ammonium herbicide tolerant LLRICE62 Crop- rice produced byinserting a modified Science phosphinothricin acetyltransferase (PAT)encoding gene from the soil bacterium Streptomyces hygroscopicus).LLRICE601 Bayer Glufosinate ammonium herbicide tolerant Crop- riceproduced by inserting a modified Science phosphinothricinacetyltransferase (PAT) (Aventis encoding gene from the soil bacteriumCrop- Streptomyces hygroscopicus). Science (AgrEvo)) PWC16 BASF Inc.Tolerance to the imidazolinone herbicide, imazethapyr, induced bychemical mutagenesis of the acetolactate synthase (ALS) enzyme usingethyl methanesulfonate (EMS).

TABLE 6F Zea mays L. Maize Event Company Description 176 SyngentaInsect-resistant maize produced by Seeds, inserting the Cry1Ab gene fromInc. Bacillus thuringiensis subsp. kurstaki. The genetic modificationaffords resistance to attack by the European corn borer (ECB). 3751IRPioneer Selection of somaclonal variants by Hi-Bred culture of embryoson imidazolinone Inter- containing media. national Inc. 676, 678,Pioneer Male-sterile and glufosinate ammonium 680 Hi-Bred herbicidetolerant maize produced by Inter- inserting genes encoding DNA adeninenational methylase and phosphinothricin Inc. acetyltransferase (PAT)from Escherichia coli and Streptomyces viridochromogenes, respectively.B16 Dekalb Glufosinate ammonium herbicide tolerant (DLL25) Geneticsmaize produced by inserting the gene Corpora- encoding phosphinothricintion acetyltransferase (PAT) from Streptomyces hygroscopicus. BT11Syngenta Insect-resistant and herbicide tolerant (X4334CBR, Seeds, maizeproduced by inserting the Cry1Ab X4734CBR) Inc. gene from Bacillusthuringiensis subsp. kurstaki, and the phosphinothricinN-acetyltransferase (PAT) encoding gene from S. viridochromogenes. BT11× Syngenta Stacked insect resistant and herbicide GA21 Seeds, tolerantmaize produced by conventional Inc. cross breeding of parental linesBT11 (OECD unique identifier: SYN-BTO11-1) and GA21 (OECD uniqueidentifier: MON-OOO21-9). BT11 × Syngenta Resistance to Coleopteranpests, MIR162 × Seeds, particularly corn rootworm pests MIR604 × Inc.(Diabrotica spp.) and several GA21 Lepidopteran pests of corn, includingEuropean corn borer (ECB, Ostrinia nubilalis), corn earworm (CEW,Helicoverpa zea), fall army worm (FAW, Spodoptera frugiperda), and blackcutworm (BCW, Agrotis ipsilon); tolerance to glyphosate andglufosinate-ammonium containing herbicides. BT11 × Syngenta Stackedinsect resistant and herbicide MIR162 Seeds, tolerant maize produced byconventional Inc. cross breeding of parental lines BT11 (OECD uniqueidentifier: SYN-BTO11-1) and MIR162 (OECD unique identifier:SYN-IR162-4). Resistance to the European Corn Borer and tolerance to theherbicide glufosinate ammonium (Liberty) is derived from BT11, whichcontains the Cry1Ab gene from Bacillus thuringiensis subsp. kurstaki,and the phosphinothricin N-acetyltransferase (PAT) encoding gene from S.viridochromogenes. Resistance to other Lepidopteran pests, including H.zea, S. frugiperda, A. ipsilon, and S. albicosta, is derived fromMIR162, which contains the vip3Aa gene from Bacillus thuringiensisstrain AB88. BT11 × Syngenta Bacillus thuringiensis Cry1Ab MIR162 ×Seeds, delta-endotoxin protein and the genetic MIR604 Inc. materialnecessary for its production (via elements of vector pZO1502) in EventBt11 corn (OECD Unique Identifier: SYN-BTO11-1) × Bacillus thuringiensisVip3Aa20 insecticidal protein and the genetic material necessary for itsproduction (via elements of vector pNOV1300) in Event MIR162 maize (OECDUnique Identifier: SYN-IR162-4) × modified Cry3A protein and the geneticmaterial necessary for its production (via elements of vector pZM26) inEvent MIR604 corn (OECD Unique Identifier: SYN-IR6O4-5). CBH-351 AventisInsect-resistant and glufosinate Crop- ammonium herbicide tolerant maizeScience developed by inserting genes encoding Cry9C protein fromBacillus thuringiensis subsp tolworthi and phosphinothricinacetyltransferase (PAT) from Streptomyces hygroscopicus. DAS-06275-8 DOWLepidopteran insect resistant and Agro- glufosinate ammoniumherbicide-tolerant Sciences maize variety produced by inserting the LLCCry1F gene from Bacillus thuringiensis var aizawai and thephosphinothricin acetyltransferase (PAT) from Streptomyceshygroscopicus. BT11 × Syngenta Stacked insect resistant and herbicideMIR604 Seeds, tolerant maize produced by conventional Inc. crossbreeding of parental lines BT11 (OECD unique identifier: SYN-BTO11-1)and MIR604 (OECD unique identifier: SYN-IR6O5-5). Resistance to theEuropean Corn Borer and tolerance to the herbicide glufosinate ammonium(Liberty) is derived from BT11, which contains the Cry1Ab gene fromBacillus thuringiensis subsp. kurstaki, and the phosphinothricinN-acetyltransferase (PAT) encoding gene from S. viridochromogenes. Cornrootworm-resistance is derived from MIR604 which contains the mCry3Agene from Bacillus thuringiensis. BT11 × Syngenta Stacked insectresistant and herbicide MIR604 × Seeds, tolerant maize produced byconventional GA21 Inc. cross breeding of parental lines BT11 (OECDunique identifier: SYN-BTO11-1), MIR604 (OECD unique identifier:SYN-IR6O5-5) and GA21 (OECD unique identifier: MON-OOO21-9). Resistanceto the European Corn Borer and tolerance to the herbicide glufosinateammonium (Liberty) is derived from BT11, which contains the Cry1Ab genefrom Bacillus thuringiensis subsp. kurstaki, and the phosphinothricinN-acetyltransferase (PAT) encoding gene from S. viridochromogenes. Cornrootworm-resistance is derived from MIR604 which contains the mCry3Agene from Bacillus thuringiensis. Tolerance to glyphosate herbicide isderived from GA21 which contains a a modified EPSPS gene from maize.DAS-59122-7 DOW Corn rootworm-resistant maize produced Agro- byinserting the Cry34Ab1 and Cry35Ab1 Sciences genes from Bacillusthuringiensis LLC and strain PS149B1. The PAT encoding gene Pioneer fromStreptomyces viridochromogenes was Hi-Bred introduced as a selectablemarker. Inter- national Inc. DAS-59122-7 × DOW Stacked insect resistantand herbicide TC1507 × Agro- tolerant maize produced by conventionalNK603 Sciences cross breeding of parental lines LLC and DAS-59122-7(OECD unique identifier: Pioneer DAS-59122-7) and TC1507 (OECD Hi-Bredunique identifier: DAS-O15O7-1) with Inter- NK603 (OECD uniqueidentifier: national MON-OO6O3-6). Corn rootworm- Inc. resistance isderived from DAS-59122-7 which contains the Cry34Ab1 and Cry35Ab1 genesfrom Bacillus thuringiensis strain PS149B1. Lepidopteran resistance andtolerance to glufosinate ammonium herbicide is derived from TC1507.Tolerance to glyphosate herbicide is derived from NK603. DBT418 DekalbInsect-resistant and glufosinate Genetics ammonium herbicide tolerantmaize Corpora- developed by inserting genes encoding tion Cry1AC proteinfrom Bacillus thuringiensis subsp kurstaki and phosphinothricinacetyltransferase (PAT) from Streptomyces hygroscopicus MIR604 ×Syngenta Stacked insect resistant and herbicide GA21 Seeds, tolerantmaize produced by conventional Inc. cross breeding of parental linesMIR604 (OECD unique identifier: SYN-IR6O5-5) and GA21 (OECD uniqueidentifier: MON-OOO21-9). Corn rootworm- resistance is derived fromMIR604 which contains the mCry3A gene from Bacillus thuringiensis.Tolerance to glyphosate herbicide is derived from GA21. MON80100Monsanto Insect-resistant maize produced by Company inserting the Cry1Abgene from Bacillus thuringiensis subsp. kurstaki. The geneticmodification affords resistance to attack by the European corn borer(ECB). MON802 Monsanto Insect-resistant and glyphosate Company herbicidetolerant maize produced by inserting the genes encoding the Cry1Abprotein from Bacillus thuringiensis and the5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from A. tumefaciensstrain CP4. MON809 Pioneer Resistance to European corn borer Hi-Bred(Ostrinia nubilalis) by introduction of Inter- a synthetic Cry1Ab gene.Glyphosate national resistance via introduction of the Inc. bacterialversion of a plant enzyme, 5-enolpyruvyl shikimate-3-phosphate synthase(EPSPS). MON810 Monsanto Insect-resistant maize produced by Companyinserting a truncated form of the Cry1Ab gene from Bacillusthuringiensis subsp. kurstaki HD-1. The genetic modification affordsresistance to attack by the European corn borer (ECB). MON810 × MonsantoStacked insect resistant and enhanced LY038 Company lysine content maizederived from conventional cross-breeding of the parental lines MON810(OECD identifier: MON-OO81O-6) and LY038 (OECD identifier: REN-OOO38-3).MON810 × Monsanto Stacked insect resistant and glyphosate MON88017Company tolerant maize derived from conventional cross-breeding of theparental lines MON810 (OECD identifier: MON-OO81O-6) and MON88017 (OECDidentifier: MON-88O17-3). European corn borer (ECB) resistance isderived from a truncated form of the Cry1Ab gene from Bacillusthuringiensis subsp. kurstaki HD-1 present in MON810. Corn rootwormresistance is derived from the Cry3Bb1 gene from Bacillus thuringiensissubspecies kumamotoensis strain EG4691 present in MON88017. Glyphosatetolerance is derived from a 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) encoding gene from Agrobacterium tumefaciens strain CP4 presentin MON88017. MON832 Monsanto Introduction, by particle bombardment,Company of glyphosate oxidase (GOX) and a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), an enzyme involved in theshikimate biochemical pathway for the production of the aromatic aminoacids. MON863 Monsanto Corn rootworm resistant maize produced Company byinserting the Cry3Bb1 gene from Bacillus thuringiensis subsp.kumamotoensis. MON863 × Monsanto Stacked insect resistant corn hybridMON810 Company derived from conventional cross-breeding of the parentallines MON863 (OECD identifier: MON-OO863-5) and MON810 (OECD identifier:MON-OO81O-6) MON863 × Monsanto Stacked insect resistant and herbicideMON810 × Company tolerant corn hybrid derived from NK603 conventionalcross-breeding of the stacked hybrid MON-OO863-5 × MON-OO81O-6 and NK603(OECD identifier: MON-OO6O3-6). MON863 × Monsanto Stacked insectresistant and herbicide NK603 Company tolerant corn hybrid derived fromconventional cross-breeding of the parental lines MON863 (OECDidentifier: MON-OO863-5) and NK603 (OECD identifier: MON-OO6O3-6).MON87460 Monsanto MON 87460 was developed to provide Company reducedyield loss under water-limited conditions compared to conventionalmaize. Efficacy in MON 87460 is derived by expression of the insertedBacillus subtilis cold shock protein B (CspB). MON88017 Monsanto Cornrootworm-resistant maize produced Company by inserting the Cry3Bb1 genefrom Bacillus thuringiensis subspecies kumamotoensis strain EG4691.Glyphosate tolerance derived by inserting a5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) encoding gene fromAgrobacterium tumefaciens strain CP4. MON89034 Monsanto Maize eventexpressing two different Company insecticidal proteins from Bacillusthuringiensis providing resistance to number of Lepidopteran pests.MON89034 × Monsanto Stacked insect resistant and glyphosate MON88017Company tolerant maize derived from conventional cross-breeding of theparental lines MON89034 (OECD identifier: MON-89O34-3) and MON88017(OECD identifier: MON-88O17-3). Resistance to Lepidopteran insects isderived from two Cry genes present in MON89043. Corn rootworm resistanceis derived from a single Cry genes and glyphosate tolerance is derivedfrom the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) encodinggene from Agrobacterium tumefaciens present in MON88017. MON89034 ×Monsanto Stacked insect resistant and herbicide NK603 Company tolerantmaize produced by conventional cross breeding of parental lines MON89034(OECD identifier: MON-89O34-3) with NK603 (OECD unique identifier:MON-OO6O3-6). Resistance to Lepidopteran insects is derived from two Crygenes present in MON89043. Tolerance to glyphosate herbicide is derivedfrom NK603. NK603 × Monsanto Stacked insect resistant and herbicideMON810 Company tolerant corn hybrid derived from conventionalcross-breeding of the parental lines NK603 (OECD identifier:MON-OO6O3-6) and MON810 (OECD identifier: MON-OO81O-6). MON89034 ×Monsanto Stacked insect resistant and herbicide TC1507 × Companytolerant maize produced by conventional MON88017 × and cross breeding ofparental lines: DAS-59122-7 Mycogen MON89034, TC1507, MON88017, andSeeds DAS-59122. Resistance to the c/o Dow above-ground and below-groundinsect Agro- pests and tolerance to glyphosate and Sciencesglufosinate-ammonium containing LLC herbicides. MS3 Bayer Male sterilitycaused by expression of Crop- the barnase ribonuclease gene from ScienceBacillus amyloliquefaciens; PPT (Aventis resistance was via Crop-PPT-acetyltransferase (PAT). Science (AgrEvo)) MS6 Bayer Male sterilitycaused by expression of Crop- the barnase ribonuclease gene from ScienceBacillus amyloliquefaciens; PPT (Aventis resistance was via Crop-PPT-acetyltransferase (PAT). Science (AgrEvo)) NK603 MonsantoIntroduction, by particle bombardment, Company of a modified5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS), an enzyme involvedin the shikimate biochemical pathway for the production of the aromaticamino acids. NK603 × Monsanto Stacked glufosinate ammonium and T25Company glyphosate herbicide tolerant maize hybrid derived fromconventional cross- breeding of the parental lines NK603 (OECDidentifier: MON-OO6O3-6) and T25 (OECD identifier: ACS-ZM003-2). T25 ×Bayer Stacked insect resistant and herbicide MON810 Crop- tolerant cornhybrid derived from Science conventional cross-breeding of the (Aventisparental lines T25 (OECD identifier: Crop- ACS-ZMOO3-2) and MON810 (OECDScience identifier: MON-OO81O-6). (AgrEvo)) TC1507 MycogenInsect-resistant and glufosinate (c/o Dow ammonium herbicide tolerantmaize Agro- produced by inserting the Cry1F gene Sciences); fromBacillus thuringiensis var. Pioneer aizawai and the phosphinothricin(c/o N-acetyltransferase encoding gene from DuPont) Streptomycesviridochromogenes. TC1507 × DOW Stacked insect resistant and herbicideNK603 Agro- tolerant corn hybrid derived from Sciences conventionalcross-breeding of the LLC parental lines 1507 (OECD identifier:DAS-O15O7-1) and NK603 (OECD identifier: MON-OO6O3-6). TC1507 × DOWStacked insect resistant and herbicide DAS-59122-7 Agro- tolerant maizeproduced by conventional Sciences cross breeding of parental linesTC1507 LLC and (OECD unique identifier: DAS-O15O7-1) Pioneer withDAS-59122-7 (OECD unique Hi-Bred identifier: DAS-59122-7). ResistanceInter- to Lepidopteran insects is derived national from TC1507 due thepresence of the Inc. Cry1F gene from Bacillus thuringiensis var.aizawai. Corn rootworm-resistance is derived from DAS-59122-7 whichcontains the Cry34Ab1 and Cry35Ab1 genes from Bacillus thuringiensisstrain PS149B1. Tolerance to glufosinate ammonium herbicide is derivedfrom TC1507 from the phosphinothricin N-acetyltransferase encoding genefrom Streptomyces viridochromogenes.

Other events with regulatory approval are well known to one skilled inthe art and can be found at the Center for Environmental Risk Assessment(cera-gmc.org/?action=gm_crop_database, which can be accessed using thewww prefix) and at the International Service for the Acquisition ofAgri-Biotech Applications (isaaa.org/gmapprovaldatabase/default.asp,which can be accessed using the www prefix).

Gene Silencing

In some embodiments the stacked trait may be in the form of silencing ofone or more polynucleotides of interest resulting in suppression of oneor more target pest polypeptides. In some embodiments the silencing isachieved through the use of a suppression DNA construct.

In some embodiments one or more polynucleotide encoding the polypeptidesof the PtIP-50 polypeptide or the PtIP-65 polypeptide or fragments orvariants thereof may be stacked with one or more polynucleotidesencoding one or more polypeptides having insecticidal activity oragronomic traits as set forth supra and optionally may further includeone or more polynucleotides providing for gene silencing of one or moretarget polynucleotides as discussed infra.

“Suppression DNA construct” is a recombinant DNA construct which whentransformed or stably integrated into the genome of the plant, resultsin “silencing” of a target gene in the plant. The target gene may beendogenous or transgenic to the plant. “Silencing,” as used herein withrespect to the target gene, refers generally to the suppression oflevels of mRNA or protein/enzyme expressed by the target gene, and/orthe level of the enzyme activity or protein functionality. The term“suppression” includes lower, reduce, decline, decrease, inhibit,eliminate and prevent. “Silencing” or “gene silencing” does not specifymechanism and is inclusive, and not limited to, anti-sense,cosuppression, viral-suppression, hairpin suppression, stem-loopsuppression, RNAi-based approaches and small RNA-based approaches.

A suppression DNA construct may comprise a region derived from a targetgene of interest and may comprise all or part of the nucleic acidsequence of the sense strand (or antisense strand) of the target gene ofinterest. Depending upon the approach to be utilized, the region may be100% identical or less than 100% identical (e.g., at least 50% or anyinteger between 51% and 100% identical) to all or part of the sensestrand (or antisense strand) of the gene of interest.

Suppression DNA constructs are well-known in the art, are readilyconstructed once the target gene of interest is selected, and include,without limitation, cosuppression constructs, antisense constructs,viral-suppression constructs, hairpin suppression constructs, stem-loopsuppression constructs, double-stranded RNA-producing constructs, andmore generally, RNAi (RNA interference) constructs and small RNAconstructs such as siRNA (short interfering RNA) constructs and miRNA(microRNA) constructs.

“Antisense inhibition” refers to the production of antisense RNAtranscripts capable of suppressing the expression of the target protein.

“Antisense RNA” refers to an RNA transcript that is complementary to allor part of a target primary transcript or mRNA and that blocks theexpression of a target isolated nucleic acid fragment (U.S. Pat. No.5,107,065). The complementarity of an antisense RNA may be with any partof the specific gene transcript, i.e., at the 5′ non-coding sequence, 3′non-coding sequence, introns or the coding sequence.

“Cosuppression” refers to the production of sense RNA transcriptscapable of suppressing the expression of the target protein. “Sense” RNArefers to RNA transcript that includes the mRNA and can be translatedinto protein within a cell or in vitro. Cosuppression constructs inplants have been previously designed by focusing on overexpression of anucleic acid sequence having homology to a native mRNA, in the senseorientation, which results in the reduction of all RNA having homologyto the overexpressed sequence (see, Vaucheret, et al., (1998) Plant J.16:651-659 and Gura, (2000) Nature 404:804-808).

Another variation describes the use of plant viral sequences to directthe suppression of proximal mRNA encoding sequences (PCT Publication WO1998/36083).

Recent work has described the use of “hairpin” structures thatincorporate all or part, of an mRNA encoding sequence in a complementaryorientation that results in a potential “stem-loop” structure for theexpressed RNA (PCT Publication WO 1999/53050). In this case the stem isformed by polynucleotides corresponding to the gene of interest insertedin either sense or anti-sense orientation with respect to the promoterand the loop is formed by some polynucleotides of the gene of interest,which do not have a complement in the construct. This increases thefrequency of cosuppression or silencing in the recovered transgenicplants. For review of hairpin suppression, see, Wesley, et al., (2003)Methods in Molecular Biology, Plant Functional Genomics: Methods andProtocols 236:273-286.

A construct where the stem is formed by at least 30 nucleotides from agene to be suppressed and the loop is formed by a random nucleotidesequence has also effectively been used for suppression (PCT PublicationWO 1999/61632).

The use of poly-T and poly-A sequences to generate the stem in thestem-loop structure has also been described (PCT Publication WO2002/00894).

Yet another variation includes using synthetic repeats to promoteformation of a stem in the stem-loop structure. Transgenic organismsprepared with such recombinant DNA fragments have been shown to havereduced levels of the protein encoded by the nucleotide fragment formingthe loop as described in PCT Publication WO 2002/00904.

RNA interference refers to the process of sequence-specificpost-transcriptional gene silencing in animals mediated by shortinterfering RNAs (siRNAs) (Fire, et al., (1998) Nature 391:806). Thecorresponding process in plants is commonly referred to aspost-transcriptional gene silencing (PTGS) or RNA silencing and is alsoreferred to as quelling in fungi. The process of post-transcriptionalgene silencing is thought to be an evolutionarily-conserved cellulardefense mechanism used to prevent the expression of foreign genes and iscommonly shared by diverse flora and phyla (Fire, et al., (1999) TrendsGenet. 15:358). Such protection from foreign gene expression may haveevolved in response to the production of double-stranded RNAs (dsRNAs)derived from viral infection or from the random integration oftransposon elements into a host genome via a cellular response thatspecifically destroys homologous single-stranded RNA of viral genomicRNA. The presence of dsRNA in cells triggers the RNAi response through amechanism that has yet to be fully characterized.

The presence of long dsRNAs in cells stimulates the activity of aribonuclease III enzyme referred to as dicer. Dicer is involved in theprocessing of the dsRNA into short pieces of dsRNA known as shortinterfering RNAs (siRNAs) (Berstein, et al., (2001) Nature 409:363).Short interfering RNAs derived from dicer activity are typically about21 to about 23 nucleotides in length and comprise about 19 base pairduplexes (Elbashir, et al., (2001) Genes Dev. 15:188). Dicer has alsobeen implicated in the excision of 21- and 22-nucleotide small temporalRNAs (stRNAs) from precursor RNA of conserved structure that areimplicated in translational control (Hutvagner, et al., (2001) Science293:834). The RNAi response also features an endonuclease complex,commonly referred to as an RNA-induced silencing complex (RISC), whichmediates cleavage of single-stranded RNA having sequence complementarityto the antisense strand of the siRNA duplex. Cleavage of the target RNAtakes place in the middle of the region complementary to the antisensestrand of the siRNA duplex (Elbashir, et al., (2001) Genes Dev. 15:188).In addition, RNA interference can also involve small RNA (e.g., miRNA)mediated gene silencing, presumably through cellular mechanisms thatregulate chromatin structure and thereby prevent transcription of targetgene sequences (see, e.g., Allshire, (2002) Science 297:1818-1819;Volpe, et al., (2002) Science 297:1833-1837; Jenuwein, (2002) Science297:2215-2218 and Hall, et al., (2002) Science 297:2232-2237). As such,miRNA molecules of the disclosure can be used to mediate gene silencingvia interaction with RNA transcripts or alternately by interaction withparticular gene sequences, wherein such interaction results in genesilencing either at the transcriptional or post-transcriptional level.

Methods and compositions are further provided which allow for anincrease in RNAi produced from the silencing element. In suchembodiments, the methods and compositions employ a first polynucleotidecomprising a silencing element for a target pest sequence operablylinked to a promoter active in the plant cell; and, a secondpolynucleotide comprising a suppressor enhancer element comprising thetarget pest sequence or an active variant or fragment thereof operablylinked to a promoter active in the plant cell. The combined expressionof the silencing element with suppressor enhancer element leads to anincreased amplification of the inhibitory RNA produced from thesilencing element over that achievable with only the expression of thesilencing element alone. In addition to the increased amplification ofthe specific RNAi species itself, the methods and compositions furtherallow for the production of a diverse population of RNAi species thatcan enhance the effectiveness of disrupting target gene expression. Assuch, when the suppressor enhancer element is expressed in a plant cellin combination with the silencing element, the methods and compositioncan allow for the systemic production of RNAi throughout the plant; theproduction of greater amounts of RNAi than would be observed with justthe silencing element construct alone; and, the improved loading of RNAiinto the phloem of the plant, thus providing better control of phloemfeeding insects by an RNAi approach. Thus, the various methods andcompositions provide improved methods for the delivery of inhibitory RNAto the target organism. See, for example, US Patent ApplicationPublication 2009/0188008.

As used herein, a “suppressor enhancer element” comprises apolynucleotide comprising the target sequence to be suppressed or anactive fragment or variant thereof. It is recognize that the suppressorenhancer element need not be identical to the target sequence, butrather, the suppressor enhancer element can comprise a variant of thetarget sequence, so long as the suppressor enhancer element hassufficient sequence identity to the target sequence to allow for anincreased level of the RNAi produced by the silencing element over thatachievable with only the expression of the silencing element. Similarly,the suppressor enhancer element can comprise a fragment of the targetsequence, wherein the fragment is of sufficient length to allow for anincreased level of the RNAi produced by the silencing element over thatachievable with only the expression of the silencing element.

It is recognized that multiple suppressor enhancer elements from thesame target sequence or from different target sequences or fromdifferent regions of the same target sequence can be employed. Forexample, the suppressor enhancer elements employed can comprisefragments of the target sequence derived from different region of thetarget sequence (i.e., from the 3′UTR, coding sequence, intron, and/or5′UTR). Further, the suppressor enhancer element can be contained in anexpression cassette, as described elsewhere herein, and in specificembodiments, the suppressor enhancer element is on the same or on adifferent DNA vector or construct as the silencing element. Thesuppressor enhancer element can be operably linked to a promoter asdisclosed herein. It is recognized that the suppressor enhancer elementcan be expressed constitutively or alternatively, it may be produced ina stage-specific manner employing the various inducible ortissue-preferred or developmentally regulated promoters that arediscussed elsewhere herein.

In specific embodiments, employing both a silencing element and thesuppressor enhancer element the systemic production of RNAi occursthroughout the entire plant. In further embodiments, the plant or plantparts of the disclosure have an improved loading of RNAi into the phloemof the plant than would be observed with the expression of the silencingelement construct alone and, thus provide better control of phloemfeeding insects by an RNAi approach. In specific embodiments, theplants, plant parts and plant cells of the disclosure can further becharacterized as allowing for the production of a diversity of RNAispecies that can enhance the effectiveness of disrupting target geneexpression.

In specific embodiments, the combined expression of the silencingelement and the suppressor enhancer element increases the concentrationof the inhibitory RNA in the plant cell, plant, plant part, plant tissueor phloem over the level that is achieved when the silencing element isexpressed alone.

As used herein, an “increased level of inhibitory RNA” comprises anystatistically significant increase in the level of RNAi produced in aplant having the combined expression when compared to an appropriatecontrol plant. For example, an increase in the level of RNAi in theplant, plant part or the plant cell can comprise at least about a 1%,about a 1%-5%, about a 5%-10%, about a 10%-20%, about a 20%-30%, about a30%-40%, about a 40%-50%, about a 50%-60%, about 60-70%, about 70%-80%,about a 80%-90%, about a 90%-100% or greater increase in the level ofRNAi in the plant, plant part, plant cell or phloem when compared to anappropriate control. In other embodiments, the increase in the level ofRNAi in the plant, plant part, plant cell or phloem can comprise atleast about a 1 fold, about a 1 fold-5 fold, about a 5 fold-10 fold,about a 10 fold-20 fold, about a 20 fold-30 fold, about a 30 fold-40fold, about a 40 fold-50 fold, about a 50 fold-60 fold, about 60 fold-70fold, about 70 fold-80 fold, about a 80 fold-90 fold, about a 90fold-100 fold or greater increase in the level of RNAi in the plant,plant part, plant cell or phloem when compared to an appropriatecontrol. Examples of combined expression of the silencing element withsuppressor enhancer element for the control of Stinkbugs and Lygus canbe found in US Patent Application Publication 2011/0301223 and US PatentApplication Publication 2009/0192117.

Some embodiments relate to down-regulation of expression of target genesin insect pest species by interfering ribonucleic acid (RNA) molecules.PCT Publication WO 2007/074405 describes methods of inhibitingexpression of target genes in invertebrate pests including Coloradopotato beetle. PCT Publication WO 2005/110068 describes methods ofinhibiting expression of target genes in invertebrate pests including inparticular Western corn rootworm as a means to control insectinfestation. Furthermore, PCT Publication WO 2009/091864 describescompositions and methods for the suppression of target genes from insectpest species including pests from the Lygus genus. Nucleic acidmolecules including RNAi for targeting the vacuolar ATPase H subunit,useful for controlling a coleopteran pest population and infestation asdescribed in US Patent Application Publication 2012/0198586. PCTPublication WO 2012/055982 describes ribonucleic acid (RNA or doublestranded RNA) that inhibits or down regulates the expression of a targetgene that encodes: an insect ribosomal protein such as the ribosomalprotein L19, the ribosomal protein L40 or the ribosomal protein 527A; aninsect proteasome subunit such as the Rpn6 protein, the Pros 25, theRpn2 protein, the proteasome beta 1 subunit protein or the Pros beta 2protein; an insect β-coatomer of the COPI vesicle, the γ-coatomer of theCOPI vesicle, the β′-coatomer protein or the ζ-coatomer of the COPIvesicle; an insect Tetraspanine 2 A protein which is a putativetransmembrane domain protein; an insect protein belonging to the actinfamily such as Actin 5C; an insect ubiquitin-5E protein; an insect Sec23protein which is a GTPase activator involved in intracellular proteintransport; an insect crinkled protein which is an unconventional myosinwhich is involved in motor activity; an insect crooked neck proteinwhich is involved in the regulation of nuclear alternative mRNAsplicing; an insect vacuolar H+-ATPase G-subunit protein and an insectTbp-1 such as Tat-binding protein. US Patent Application Publications2012/029750, US 20120297501, and 2012/0322660 describe interferingribonucleic acids (RNA or double stranded RNA) that functions uponuptake by an insect pest species to down-regulate expression of a targetgene in said insect pest, wherein the RNA comprises at least onesilencing element wherein the silencing element is a region ofdouble-stranded RNA comprising annealed complementary strands, onestrand of which comprises or consists of a sequence of nucleotides whichis at least partially complementary to a target nucleotide sequencewithin the target gene. US Patent Application Publication 2012/0164205describe potential targets for interfering double stranded ribonucleicacids for inhibiting invertebrate pests including: a Chd3 HomologousSequence, a Beta-Tubulin Homologous Sequence, a 40 kDa V-ATPaseHomologous Sequence, a EF1a Homologous Sequence, a 26S ProteosomeSubunit p28 Homologous Sequence, a Juvenile Hormone Epoxide HydrolaseHomologous Sequence, a Swelling Dependent Chloride Channel ProteinHomologous Sequence, a Glucose-6-Phosphate 1-Dehydrogenase ProteinHomologous Sequence, an Act42A Protein Homologous Sequence, aADP-Ribosylation Factor 1 Homologous Sequence, a Transcription FactorIIB Protein Homologous Sequence, a Chitinase Homologous Sequences, aUbiquitin Conjugating Enzyme Homologous Sequence, aGlyceraldehyde-3-Phosphate Dehydrogenase Homologous Sequence, anUbiquitin B Homologous Sequence, a Juvenile Hormone Esterase Homolog,and an Alpha Tubuliln Homologous Sequence.

Use in Pesticidal Control

General methods for employing strains comprising a nucleic acid sequenceof the embodiments or a variant thereof, in pesticide control or inengineering other organisms as pesticidal agents are known in the art.See, for example U.S. Pat. No. 5,039,523 and EP 0480762A2.

Microorganism hosts that are known to occupy the “phytosphere”(phylloplane, phyllosphere, rhizosphere, and/or rhizoplana) of one ormore crops of interest may be selected. These microorganisms areselected so as to be capable of successfully competing in the particularenvironment with the wild-type microorganisms, provide for stablemaintenance and expression of the gene expressing the PtIP-50polypeptide or the PtIP-65 polypeptide, and desirably, provide forimproved protection of the pesticide from environmental degradation andinactivation.

Such microorganisms include bacteria, algae, and fungi. Of particularinterest are microorganisms such as bacteria, e.g., Pseudomonas,Erwinia, Serratia, Klebsiella, Xanthomonas, Streptomyces, Rhizobium,Rhodopseudomonas, Methylius, Agrobacterium, Acetobacter, Lactobacillus,Arthrobacter, Azotobacter, Leuconostoc, and Alcaligenes, fungi,particularly yeast, e.g., Saccharomyces, Cryptococcus, Kluyveromyces,Sporobolomyces, Rhodotorula, and Aureobasidium. Of particular interestare such phytosphere bacterial species as Pseudomonas syringae,Pseudomonas fluorescens, Pseudomonas chlororaphis, Serratia marcescens,Acetobacter xylinum, Agrobacteria, Rhodopseudomonas spheroides,Xanthomonas campestris, Rhizobium melioti, Alcaligenes entrophus,Clavibacter xyli and Azotobacter vinelandii and phytosphere yeastspecies such as Rhodotorula rubra, R. glutinis, R. marina, R.aurantiaca, Cryptococcus albidus, C. diffluens, C. laurentii,Saccharomyces rosei, S. pretoriensis, S. cerevisiae, Sporobolomycesroseus, S. odorus, Kluyveromyces veronae, and Aureobasidium pollulans.Of particular interest are the pigmented microorganisms. Host organismsof particular interest include yeast, such as Rhodotorula spp.,Aureobasidium spp., Saccharomyces spp. (such as S. cerevisiae),Sporobolomyces spp., phylloplane organisms such as Pseudomonas spp.(such as P. aeruginosa, P. fluorescens, P. chlororaphis), Erwinia spp.,and Flavobacterium spp., and other such organisms, includingAgrobacterium tumefaciens, E. coli, Bacillus subtilis, Bacillus cereusand the like.

Genes encoding the PtIP-50 polypeptide and/or the PtIP-65 polypeptide ofthe embodiments can be introduced into microorganisms that multiply onplants (epiphytes) to deliver PtIP-50 polypeptide and/or the PtIP-65polypeptides to potential target pests. Epiphytes, for example, can begram-positive or gram-negative bacteria.

Root-colonizing bacteria, for example, can be isolated from the plant ofinterest by methods known in the art. Specifically, a Bacillus cereusstrain that colonizes roots can be isolated from roots of a plant (see,for example, Handelsman et al. (1991) Appl. Environ. Microbiol.56:713-718). Genes encoding the PtIP-50 polypeptide and/or the PtIP-65polypeptide of the embodiments can be introduced into a root-colonizingBacillus cereus by standard methods known in the art.

Genes encoding PtIP-50 polypeptides and/or the PtIP-65 polypeptides canbe introduced, for example, into the root-colonizing Bacillus by meansof electro transformation. Specifically, genes encoding the PtIP-50polypeptides and/or the PtIP-65 polypeptides can be cloned into ashuttle vector, for example, pHT3101 (Lerecius, et al., (1989) FEMSMicrobiol. Letts. 60:211-218. The shuttle vector pHT3101 containing thecoding sequence for the particular PtIP-50 polypeptide and/or thePtIP-65 polypeptide gene can, for example, be transformed into theroot-colonizing Bacillus by means of electroporation (Lerecius, et al.,(1989) FEMS Microbiol. Letts. 60:211-218).

Expression systems can be designed so that PtIP-50 polypeptides and/orthe PtIP-65 polypeptides are secreted outside the cytoplasm ofgram-negative bacteria, such as E. coli, for example. Advantages ofhaving a PtIP-50 polypeptide and/or a PtIP-65 polypeptide secreted are:(1) avoidance of potential cytotoxic effects of the PtIP-50 polypeptideand/or the PtIP-65 polypeptide expressed; and (2) improvement in theefficiency of purification of the PtIP-50 polypeptide and/or the PtIP-65polypeptide, including, but not limited to, increased efficiency in therecovery and purification of the protein per volume cell broth anddecreased time and/or costs of recovery and purification per unitprotein.

PtIP-50 polypeptides and/or the PtIP-65 polypeptides can be made to besecreted in E. coli, for example, by fusing an appropriate E. colisignal peptide to the amino-terminal end of the PtIP-50 polypeptideand/or the PtIP-65 polypeptide. Signal peptides recognized by E. colican be found in proteins already known to be secreted in E. coli, forexample the OmpA protein (Ghrayeb, et al., (1984) EMBO J, 3:2437-2442).OmpA is a major protein of the E. coli outer membrane, and thus itssignal peptide is thought to be efficient in the translocation process.Also, the OmpA signal peptide does not need to be modified beforeprocessing as may be the case for other signal peptides, for examplelipoprotein signal peptide (Duffaud, et al., (1987) Meth. Enzymol.153:492).

PtIP-50 polypeptides and/or the PtIP-65 polypeptides of the embodimentscan be fermented in a bacterial host and the resulting bacteriaprocessed and used as a microbial spray in the same manner that Btstrains have been used as insecticidal sprays. In the case of a PtIP-50polypeptide(s) and/or the PtIP-65 polypeptide(s) that is secreted fromBacillus, the secretion signal is removed or mutated using proceduresknown in the art. Such mutations and/or deletions prevent secretion ofthe PtIP-50 polypeptide(s) and/or the PtIP-65 polypeptide(s) into thegrowth medium during the fermentation process. The PtIP-50 polypeptideand/or the PtIP-65 polypeptide are retained within the cell, and thecells are then processed to yield the encapsulated PtIP-50 polypeptideand/or the PtIP-65 polypeptide. Any suitable microorganism can be usedfor this purpose. Pseudomonas has been used to express Bt toxins asencapsulated proteins and the resulting cells processed and sprayed asan insecticide (Gaertner, et al., (1993), in: Advanced EngineeredPesticides, ed. Kim).

Alternatively, the PtIP-50 polypeptide and/or the PtIP-65 polypeptideare produced by introducing a heterologous gene into a cellular host.Expression of the heterologous gene results, directly or indirectly, inthe intracellular production and maintenance of the pesticide. Thesecells are then treated under conditions that prolong the activity of thetoxin produced in the cell when the cell is applied to the environmentof target pest(s). The resulting product retains the toxicity of thetoxin. These naturally encapsulated PtIP-50 polypeptide and/or thePtIP-65 polypeptide may then be formulated in accordance withconventional techniques for application to the environment hosting atarget pest, e.g., soil, water, and foliage of plants. See, for exampleEPA 0192319, and the references cited therein.

Pesticidal Compositions

In some embodiments the active ingredients can be applied in the form ofcompositions and can be applied to the crop area or plant to be treated,simultaneously or in succession, with other compounds. These compoundscan be fertilizers, weed killers, Cryoprotectants, surfactants,detergents, pesticidal soaps, dormant oils, polymers, and/ortime-release or biodegradable carrier formulations that permit long-termdosing of a target area following a single application of theformulation. They can also be selective herbicides, chemicalinsecticides, virucides, microbicides, amoebicides, pesticides,fungicides, bacteriocides, nematocides, molluscicides or mixtures ofseveral of these preparations, if desired, together with furtheragriculturally acceptable carriers, surfactants or application-promotingadjuvants customarily employed in the art of formulation. Suitablecarriers and adjuvants can be solid or liquid and correspond to thesubstances ordinarily employed in formulation technology, e.g. naturalor regenerated mineral substances, solvents, dispersants, wettingagents, tackifiers, binders or fertilizers. Likewise the formulationsmay be prepared into edible “baits” or fashioned into pest “traps” topermit feeding or ingestion by a target pest of the pesticidalformulation.

Methods of applying an active ingredient or an agrochemical compositionthat contains at least one of the PtIP-50 polypeptide and/or the PtIP-65polypeptide produced by the bacterial strains include leaf application,seed coating and soil application. The number of applications and therate of application depend on the intensity of infestation by thecorresponding pest.

The composition may be formulated as a powder, dust, pellet, granule,spray, emulsion, colloid, solution or such like, and may be prepared bysuch conventional means as desiccation, lyophilization, homogenation,extraction, filtration, centrifugation, sedimentation or concentrationof a culture of cells comprising the polypeptide. In all suchcompositions that contain at least one such pesticidal polypeptide, thepolypeptide may be present in a concentration of from about 1% to about99% by weight.

Lepidopteran, Dipteran, Heteropteran, nematode, Hemiptera or Coleopteranpests may be killed or reduced in numbers in a given area by the methodsof the disclosure or may be prophylactically applied to an environmentalarea to prevent infestation by a susceptible pest. Preferably the pestingests or is contacted with, a pesticidally-effective amount of thepolypeptide. “Pesticidally-effective amount” as used herein refers to anamount of the pesticide that is able to bring about death to at leastone pest or to noticeably reduce pest growth, feeding or normalphysiological development. This amount will vary depending on suchfactors as, for example, the specific target pests to be controlled, thespecific environment, location, plant, crop or agricultural site to betreated, the environmental conditions and the method, rate,concentration, stability, and quantity of application of thepesticidally-effective polypeptide composition. The formulations mayalso vary with respect to climatic conditions, environmentalconsiderations, and/or frequency of application and/or severity of pestinfestation.

The pesticide compositions described may be made by formulating eitherthe bacterial cell, Crystal and/or spore suspension or isolated proteincomponent with the desired agriculturally-acceptable carrier. Thecompositions may be formulated prior to administration in an appropriatemeans such as lyophilized, freeze-dried, desiccated or in an aqueouscarrier, medium or suitable diluent, such as saline or other buffer. Theformulated compositions may be in the form of a dust or granularmaterial or a suspension in oil (vegetable or mineral) or water oroil/water emulsions or as a wettable powder or in combination with anyother carrier material suitable for agricultural application. Suitableagricultural carriers can be solid or liquid and are well known in theart. The term “agriculturally-acceptable carrier” covers all adjuvants,inert components, dispersants, surfactants, tackifiers, binders, etc.that are ordinarily used in pesticide formulation technology; these arewell known to those skilled in pesticide formulation. The formulationsmay be mixed with one or more solid or liquid adjuvants and prepared byvarious means, e.g., by homogeneously mixing, blending and/or grindingthe pesticidal composition with suitable adjuvants using conventionalformulation techniques. Suitable formulations and application methodsare described in U.S. Pat. No. 6,468,523, herein incorporated byreference. The plants can also be treated with one or more chemicalcompositions, including one or more herbicide, insecticides orfungicides. Exemplary chemical compositions include: Fruits/VegetablesHerbicides: Atrazine, Bromacil, Diuron, Glyphosate, Linuron, Metribuzin,Simazine, Trifluralin, Fluazifop, Glufosinate, Halo sulfuron Gowan,Paraquat, Propyzamide, Sethoxydim, Butafenacil, Halosulfuron,Indaziflam; Fruits/Vegetables Insecticides: Aldicarb, Bacillusthuriengiensis, Carbaryl, Carbofuran, Chlorpyrifos, Cypermethrin,Deltamethrin, Diazinon, Malathion, Abamectin,Cyfluthrin/beta-cyfluthrin, Esfenvalerate, Lambda-cyhalothrin,Acequinocyl, Bifenazate, Methoxyfenozide, Novaluron, Chromafenozide,Thiacloprid, Dinotefuran, FluaCrypyrim, Tolfenpyrad, Clothianidin,Spirodiclofen, Gamma-cyhalothrin, Spiromesifen, Spinosad, Rynaxypyr,Cyazypyr, Spinoteram, Triflumuron, Spirotetramat, Imidacloprid,Flubendiamide, Thiodicarb, Metaflumizone, Sulfoxaflor, Cyflumetofen,Cyanopyrafen, Imidacloprid, Clothianidin, Thiamethoxam, Spinotoram,Thiodicarb, Flonicamid, Methiocarb, Emamectin-benzoate, lndoxacarb,Forthiazate, Fenamiphos, Cadusaphos, Pyriproxifen, Fenbutatin-oxid,Hexthiazox, Methomyl,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on;Fruits/Vegetables Fungicides: Carbendazim, Chlorothalonil, EBDCs,Sulphur, Thiophanate-methyl, Azoxystrobin, Cymoxanil, Fluazinam,Fosetyl, Iprodione, Kresoxim-methyl, Metalaxyl/mefenoxam,Trifloxystrobin, Ethaboxam, Iprovalicarb, Trifloxystrobin, Fenhexamid,Oxpoconazole fumarate, Cyazofamid, Fenamidone, Zoxamide, Picoxystrobin,Pyraclostrobin, Cyflufenamid, Boscalid; Cereals Herbicides: Isoproturon,Bromoxynil, loxynil, Phenoxies, Chlorsulfuron, Clodinafop, Diclofop,Diflufenican, Fenoxaprop, Florasulam, Fluoroxypyr, Metsulfuron,Triasulfuron, Flucarbazone, lodosulfuron, Propoxycarbazone, Picolinafen,Mesosulfuron, Beflubutamid, Pinoxaden, Amidosulfuron, ThifensulfuronMethyl, Tribenuron, Flupyrsulfuron, Sulfosulfuron, Pyrasulfotole,Pyroxsulam, Flufenacet, Tralkoxydim, Pyroxasulfon; Cereals Fungicides:Carbendazim, Chlorothalonil, Azoxystrobin, Cyproconazole, Cyprodinil,Fenpropimorph, Epoxiconazole, Kresoxim-methyl, Quinoxyfen, Tebuconazole,Trifloxystrobin, Simeconazole, Picoxystrobin, Pyraclostrobin,Dimoxystrobin, Prothioconazole, Fluoxastrobin; Cereals Insecticides:Dimethoate, Lambda-cyhalthrin, Deltamethrin, alpha-Cypermethrin,β-cyfluthrin, Bifenthrin, Imidacloprid, Clothianidin, Thiamethoxam,Thiacloprid, Acetamiprid, Dinetofuran, Clorphyriphos, Metamidophos,Oxidemethon-methyl, Pirimicarb, Methiocarb; Maize Herbicides: Atrazine,Alachlor, Bromoxynil, Acetochlor, Dicamba, Clopyralid, (S-)Dimethenamid, Glufosinate, Glyphosate, Isoxaflutole, (S-) Metolachlor,Mesotrione, Nicosulfuron, Primisulfuron, Rimsulfuron, Sulcotrione,Foramsulfuron, Topramezone, Tembotrione, Saflufenacil, Thiencarbazone,Flufenacet, Pyroxasulfon; Maize Insecticides: Carbofuran, Chlorpyrifos,Bifenthrin, Fipronil, Imidacloprid, Lambda-Cyhalothrin, Tefluthrin,Terbufos, Thiamethoxam, Clothianidin, Spiromesifen, Flubendiamide,Triflumuron, Rynaxypyr, Deltamethrin, Thiodicarb, β-Cyfluthrin,Cypermethrin, Bifenthrin, Lufenuron, Triflumoron, Tefluthrin,Tebupirimphos, Ethiprole, Cyazypyr, Thiacloprid, Acetamiprid,Dinetofuran, Avermectin, Methiocarb, Spirodiclofen, Spirotetramat; MaizeFungicides: Fenitropan, Thiram, Prothioconazole, Tebuconazole,Trifloxystrobin; Rice Herbicides: Butachlor, Propanil, Azimsulfuron,Bensulfuron, Cyhalofop, Daimuron, Fentrazamide, Imazosulfuron,Mefenacet, Oxaziclomefone, Pyrazosulfuron, Pyributicarb, Quinclorac,Thiobencarb, Indanofan, Flufenacet, Fentrazamide, Halosulfuron,Oxaziclomefone, Benzobicyclon, Pyriftalid, Penoxsulam, Bispyribac,Oxadiargyl, Ethoxysulfuron, Pretilachlor, Mesotrione, Tefuryltrione,Oxadiazone, Fenoxaprop, Pyrimisulfan; Rice Insecticides: Diazinon,Fenitrothion, Fenobucarb, Monocrotophos, Benfuracarb, Buprofezin,Dinotefuran, Fipronil, Imidacloprid, Isoprocarb, Thiacloprid,Chromafenozide, Thiacloprid, Dinotefuran, Clothianidin, Ethiprole,Flubendiamide, Rynaxypyr, Deltamethrin, Acetamiprid, Thiamethoxam,Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Cypermethrin,Chlorpyriphos, Cartap, Methamidophos, Etofenprox, Triazophos,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Carbofuran, Benfuracarb; Rice Fungicides: Thiophanate-methyl,Azoxystrobin, Carpropamid, Edifenphos, Ferimzone, Iprobenfos,Isoprothiolane, Pencycuron, Probenazole, Pyroquilon, Tricyclazole,Trifloxystrobin, Diclocymet, Fenoxanil, Simeconazole, Tiadinil; CottonHerbicides: Diuron, Fluometuron, MSMA, Oxyfluorfen, Prometryn,Trifluralin, Carfentrazone, Clethodim, Fluazifop-butyl, Glyphosate,Norflurazon, Pendimethalin, Pyrithiobac-sodium, Trifloxysulfuron,Tepraloxydim, Glufosinate, Flumioxazin, Thidiazuron; CottonInsecticides: Acephate, Aldicarb, Chlorpyrifos, Cypermethrin,Deltamethrin, Malathion, Monocrotophos, Abamectin, Acetamiprid,Emamectin Benzoate, Imidacloprid, Indoxacarb, Lambda-Cyhalothrin,Spinosad, Thiodicarb, Gamma-Cyhalothrin, Spiromesifen, Pyridalyl,Flonicamid, Flubendiamide, Triflumuron, Rynaxypyr, Beta-Cyfluthrin,Spirotetramat, Clothianidin, Thiamethoxam, Thiacloprid, Dinetofuran,Flubendiamide, Cyazypyr, Spinosad, Spinotoram, gamma Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Thiodicarb, Avermectin, Flonicamid, Pyridalyl, Spiromesifen,Sulfoxaflor, Profenophos, Thriazophos, Endosulfan; Cotton Fungicides:Etridiazole, Metalaxyl, Quintozene; Soybean Herbicides: Alachlor,Bentazone, Trifluralin, Chlorimuron-Ethyl, Cloransulam-Methyl,Fenoxaprop, Fomesafen, Fluazifop, Glyphosate, Imazamox, Imazaquin,Imazethapyr, (S-) Metolachlor, Metribuzin, Pendimethalin, Tepraloxydim,Glufosinate; Soybean Insecticides: Lambda-cyhalothrin, Methomyl,Parathion, Thiocarb, Imidacloprid, Clothianidin, Thiamethoxam,Thiacloprid, Acetamiprid, Dinetofuran, Flubendiamide, Rynaxypyr,Cyazypyr, Spinosad, Spinotoram, Emamectin-Benzoate, Fipronil, Ethiprole,Deltamethrin, β-Cyfluthrin, gamma and lambda Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Spirotetramat, Spinodiclofen, Triflumuron, Flonicamid, Thiodicarb,beta-Cyfluthrin; Soybean Fungicides: Azoxystrobin, Cyproconazole,Epoxiconazole, Flutriafol, Pyraclostrobin, Tebuconazole,Trifloxystrobin, Prothioconazole, Tetraconazole; Sugarbeet Herbicides:Chloridazon, Desmedipham, Ethofumesate, Phenmedipham, Triallate,Clopyralid, Fluazifop, Lenacil, Metamitron, Quinmerac, Cycloxydim,Triflusulfuron, Tepraloxydim, Quizalofop; Sugarbeet Insecticides:Imidacloprid, Clothianidin, Thiamethoxam, Thiacloprid, Acetamiprid,Dinetofuran, Deltamethrin, β-Cyfluthrin, gamma/lambda Cyhalothrin,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on,Tefluthrin, Rynaxypyr, Cyaxypyr, Fipronil, Carbofuran; CanolaHerbicides: Clopyralid, Diclofop, Fluazifop, Glufosinate, Glyphosate,Metazachlor, Trifluralin Ethametsulfuron, Quinmerac, Quizalofop,Clethodim, Tepraloxydim; Canola Fungicides: Azoxystrobin, Carbendazim,Fludioxonil, Iprodione, Prochloraz, Vinclozolin; Canola Insecticides:Carbofuran organophosphates, Pyrethroids, Thiacloprid, Deltamethrin,Imidacloprid, Clothianidin, Thiamethoxam, Acetamiprid, Dinetofuran,β-Cyfluthrin, gamma and lambda Cyhalothrin, tau-Fluvaleriate, Ethiprole,Spinosad, Spinotoram, Flubendiamide, Rynaxypyr, Cyazypyr,4-[[(6-Chlorpyridin-3-yl)methyl](2,2-difluorethyl)amino]furan-2(5H)-on.

In some embodiments the herbicide is Atrazine, Bromacil, Diuron,Chlorsulfuron, Metsulfuron, Thifensulfuron Methyl, Tribenuron,Acetochlor, Dicamba, Isoxaflutole, Nicosulfuron, Rimsulfuron,Pyrithiobac-sodium, Flumioxazin, Chlorimuron-Ethyl, Metribuzin,Quizalofop, S-metolachlor, Hexazinne or combinations thereof.

In some embodiments the insecticide is Esfenvalerate,Chlorantraniliprole, Methomyl, lndoxacarb, Oxamyl or combinationsthereof.

Pesticidal and Insecticidal Activity

“Pest” includes but is not limited to, insects, fungi, bacteria,nematodes, mites, ticks and the like. Insect pests include insectsselected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera,Mallophaga, Homoptera, Hemiptera Orthroptera, Thysanoptera, Dermaptera,Isoptera, Anoplura, Siphonaptera, Trichoptera, etc., particularlyLepidoptera and Coleoptera.

Those skilled in the art will recognize that not all compounds areequally effective against all pests. Compounds of the embodimentsdisplay activity against insect pests, which may include economicallyimportant agronomic, forest, greenhouse, nursery ornamentals, food andfiber, public and animal health, domestic and commercial structure,household and stored product pests.

Larvae of the order Lepidoptera include, but are not limited to,armyworms, cutworms, loopers and heliothines in the family NoctuidaeSpodoptera frugiperda J E Smith (fall armyworm); S. exigua Hübner (beetarmyworm); S. litura Fabricius (tobacco cutworm, cluster caterpillar);Mamestra configurata Walker (bertha armyworm); M. brassicae Linnaeus(cabbage moth); Agrotis ipsilon Hufnagel (black cutworm); A. orthogoniaMorrison (western cutworm); A. subterranea Fabricius (granulatecutworm); Alabama argillacea Hübner (cotton leaf worm); Trichoplusia niHübner (cabbage looper); Pseudoplusia includens Walker (soybean looper);Anticarsia gemmatalis Hübner (velvetbean caterpillar); Hypena scabraFabricius (green cloverworm); Heliothis virescens Fabricius (tobaccobudworm); Pseudaletia unipuncta Haworth (armyworm); Athetis mindaraBarnes and Mcdunnough (rough skinned cutworm); Euxoa messoria Harris(darksided cutworm); Earias insulana Boisduval (spiny bollworm); E.vittella Fabricius (spotted bollworm); Helicoverpa armigera Hübner(American bollworm); H. zea Boddie (corn earworm or cotton bollworm);Melanchra picta Harris (zebra caterpillar); Egira (Xylomyges) curialisGrote (citrus cutworm); borers, casebearers, webworms, coneworms, andskeletonizers from the family Pyralidae Ostrinia nubilalis Hübner(European corn borer); Amyelois transitella Walker (naval orangeworm);Anagasta kuehniella Zeller (Mediterranean flour moth); Cadra cautellaWalker (almond moth); Chilo suppressalis Walker (rice stem borer); C.partellus, (sorghum borer); Corcyra cephalonica Stainton (rice moth);Crambus caliginosellus Clemens (corn root webworm); C. teterrellusZincken (bluegrass webworm); Cnaphalocrocis medinalis Guenee (rice leafroller); Desmia funeralis Hübner (grape leaffolder); Diaphania hyalinataLinnaeus (melon worm); D. nitidalis Stoll (pickleworm); Diatraeagrandiosella Dyar (southwestern corn borer), D. saccharalis Fabricius(surgarcane borer); Eoreuma loftini Dyar (Mexican rice borer); Ephestiaelutella Hübner (tobacco (cacao) moth); Galleria mellonella Linnaeus(greater wax moth); Herpetogramma licarsisalis Walker (sod webworm);Homoeosoma electellum Hulst (sunflower moth); Elasmopalpus lignosellusZeller (lesser cornstalk borer); Achroia grisella Fabricius (lesser waxmoth); Loxostege sticticalis Linnaeus (beet webworm); Orthaga thyrisalisWalker (tea tree web moth); Maruca testulalis Geyer (bean pod borer);Plodia interpunctella Hübner (Indian meal moth); Scirpophaga incertulasWalker (yellow stem borer); Udea rubigalis Guenee (celery leaftier); andleafrollers, budworms, seed worms and fruit worms in the familyTortricidae Acleris gloverana Walsingham (Western blackheaded budworm);A. variana Fernald (Eastern blackheaded budworm); Archips argyrospilaWalker (fruit tree leaf roller); A. rosana Linnaeus (European leafroller); and other Archips species, Adoxophyes orana Fischer vonRosslerstamm (summer fruit tortrix moth); Cochylis hospes Walsingham(banded sunflower moth); Cydia latiferreana Walsingham (filbertworm); C.pomonella Linnaeus (coding moth); Platynota flavedana Clemens(variegated leafroller); P. stultana Walsingham (omnivorous leafroller);Lobesia botrana Denis & Schiffermuller (European grape vine moth);Spilonota ocellana Denis & Schiffermuller (eyespotted bud moth);Endopiza viteana Clemens (grape berry moth); Eupoecilia ambiguellaHübner (vine moth); Bonagota salubricola Meyrick (Brazilian appleleafroller); Grapholita molesta Busck (oriental fruit moth); Suleimahelianthana Riley (sunflower bud moth); Argyrotaenia spp.; Choristoneuraspp.

Selected other agronomic pests in the order Lepidoptera include, but arenot limited to, Alsophila pometaria Harris (fall cankerworm); Anarsialineatella Zeller (peach twig borer); Anisota senatoria J. E. Smith(orange striped oakworm); Antheraea pernyi Gurin-Meneville (Chinese OakTussah Moth); Bombyx mori Linnaeus (Silkworm); Bucculatrix thurberiellaBusck (cotton leaf perforator); Collas eurytheme Boisduval (alfalfacaterpillar); Datana integerrima Grote & Robinson (walnut caterpillar);Dendrolimus sibiricus Tschetwerikov (Siberian silk moth), Ennomossubsignaria Hübner (elm spanworm); Erannis tiliaria Harris (lindenlooper); Euproctis chrysorrhoea Linnaeus (browntail moth); Harrisinaamericana Gurin-Meneville (grapeleaf skeletonizer); Hemileuca oliviaeCockrell (range caterpillar); Hyphantria cunea Drury (fall webworm);Keiferia lycopersicella Walsingham (tomato pinworm); Lambdinafiscellaria fiscellaria Hulst (Eastern hemlock looper); L. fiscellarialugubrosa Hulst (Western hemlock looper); Leucoma salicis Linnaeus(satin moth); Lymantria dispar Linnaeus (gypsy moth); Manducaquinquemaculata Haworth (five spotted hawk moth, tomato hornworm); M.sexta Haworth (tomato hornworm, tobacco hornworm); Operophtera brumataLinnaeus (winter moth); Paleacrita vernata Peck (spring cankerworm);Papilio cresphontes Cramer (giant swallowtail orange dog); Phtyganidiacalifornica Packard (California oakworm); Phyllocnistis citrellaStainton (citrus leafminer); Phyllonotycter blancardella Fabricius(spotted tentiform leafminer); Pieris brassicae Linnaeus (large whitebutterfly); P. rapae Linnaeus (small white butterfly); P. napi Linnaeus(green veined white butterfly); Platyptilia carduidactyla Riley(artichoke plume moth); Plutella xylostella Linnaeus (diamondback moth);Pectinophora gossypiella Saunders (pink bollworm); Pontia protodiceBoisduval and Leconte (Southern cabbageworm); Sabulodes aegrotata Guenee(omnivorous looper); Schizura concinna J. E. Smith (red humpedcaterpillar); Sitotroga cerealella Olivier (Angoumois grain moth);Thaumetopoea pityocampa Schiffermuller (pine processionary caterpillar);Tineola bisselliella Hummel (webbing clothesmoth); Tuta absoluta Meyrick(tomato leafminer); Yponomeuta padella Linnaeus (ermine moth); Heliothissubflexa Guenee; Malacosoma spp. and Orgyia spp.

Of interest are larvae and adults of the order Coleoptera includingweevils from the families Anthribidae, Bruchidae and Curculionidae(including, but not limited to: Anthonomus grandis Boheman (bollweevil); Lissorhoptrus oryzophilus Kuschel (rice water weevil);Sitophilus granarius Linnaeus (granary weevil); S. oryzae Linnaeus (riceweevil); Hypera punctata Fabricius (clover leaf weevil);Cylindrocopturus adspersus LeConte (sunflower stem weevil); Smicronyxfulvus LeConte (red sunflower seed weevil); S. sordidus LeConte (graysunflower seed weevil); Sphenophorus maidis Chittenden (maize billbug));flea beetles, cucumber beetles, rootworms, leaf beetles, potato beetlesand leafminers in the family Chrysomelidae (including, but not limitedto: Leptinotarsa decemlineata Say (Colorado potato beetle); Diabroticavirgifera virgifera LeConte (western corn rootworm); D. barberi Smithand Lawrence (northern corn rootworm); D. undecimpunctata howardi Barber(southern corn rootworm); Chaetocnema pulicaria Melsheimer (corn fleabeetle); Phyllotreta cruciferae Goeze (Crucifer flea beetle);Phyllotreta striolata (stripped flea beetle); Colaspis brunnea Fabricius(grape colaspis); Oulema melanopus Linnaeus (cereal leaf beetle);Zygogramma exclamationis Fabricius (sunflower beetle)); beetles from thefamily Coccinellidae (including, but not limited to: Epilachnavarivestis Mulsant (Mexican bean beetle)); chafers and other beetlesfrom the family Scarabaeidae (including, but not limited to: Popilliajaponica Newman (Japanese beetle); Cyclocephala borealis Arrow (northernmasked chafer, white grub); C. immaculata Olivier (southern maskedchafer, white grub); Rhizotrogus majalis Razoumowsky (European chafer);Phyllophaga crinita Burmeister (white grub); Ligyrus gibbosus De Geer(carrot beetle)); carpet beetles from the family Dermestidae; wirewormsfrom the family Elateridae, Eleodes spp., Melanotus spp.; Conoderusspp.; Limonius spp.; Agriotes spp.; Ctenicera spp.; Aeolus spp.; barkbeetles from the family Scolytidae and beetles from the familyTenebrionidae.

Adults and immatures of the order Diptera are of interest, includingleafminers Agromyza parvicornis Loew (corn blotch leafminer); midges(including, but not limited to: Contarinia sorghicola Coquillett(sorghum midge); Mayetiola destructor Say (Hessian fly); Sitodiplosismosellana Gehin (wheat midge); Neolasioptera murtfeldtiana Felt,(sunflower seed midge)); fruit flies (Tephritidae), Oscinella fritLinnaeus (fruit flies); maggots (including, but not limited to: Deliaplatura Meigen (seedcorn maggot); D. coarctata Fallen (wheat bulb fly)and other Delia spp., Meromyza americana Fitch (wheat stem maggot);Musca domestica Linnaeus (house flies); Fannia canicularis Linnaeus, F.femoralis Stein (lesser house flies); Stomoxys calcitrans Linnaeus(stable flies)); face flies, horn flies, blow flies, Chrysomya spp.;Phormia spp. and other muscoid fly pests, horse flies Tabanus spp.; botflies Gastrophilus spp.; Oestrus spp.; cattle grubs Hypoderma spp.; deerflies Chrysops spp.; Melophagus ovinus Linnaeus (keds) and otherBrachycera, mosquitoes Aedes spp.; Anopheles spp.; Culex spp.; blackflies Prosimulium spp.; Simulium spp.; biting midges, sand flies,sciarids, and other Nematocera.

Included as insects of interest are adults and nymphs of the ordersHemiptera and Homoptera such as, but not limited to, adelgids from thefamily Adelgidae, plant bugs from the family Miridae, cicadas from thefamily Cicadidae, leafhoppers, Empoasca spp.; from the familyCicadellidae, planthoppers from the families Cixiidae, Flatidae,Fulgoroidea, lssidae and Delphacidae, treehoppers from the familyMembracidae, psyllids from the family Psyllidae, whiteflies from thefamily Aleyrodidae, aphids from the family Aphididae, phylloxera fromthe family Phylloxeridae, mealybugs from the family Pseudococcidae,scales from the families Asterolecanidae, Coccidae, Dactylopiidae,Diaspididae, Eriococcidae Ortheziidae, Phoenicococcidae andMargarodidae, lace bugs from the family Tingidae, stink bugs from thefamily Pentatomidae, cinch bugs, Blissus spp.; and other seed bugs fromthe family Lygaeidae, spittlebugs from the family Cercopidae squash bugsfrom the family Coreidae and red bugs and cotton stainers from thefamily Pyrrhocoridae.

Agronomically important members from the order Homoptera furtherinclude, but are not limited to: Acyrthisiphon pisum Harris (pea aphid);Aphis craccivora Koch (cowpea aphid); A. fabae Scopoli (black beanaphid); A. gossypii Glover (cotton aphid, melon aphid); A. maidiradicisForbes (corn root aphid); A. pomi De Geer (apple aphid); A. spiraecolaPatch (spirea aphid); Aulacorthum solani Kaltenbach (foxglove aphid);Chaetosiphon fragaefolii Cockerell (strawberry aphid); Diuraphis noxiaKurdjumov/Mordvilko (Russian wheat aphid); Dysaphis plantagineaPaaserini (rosy apple aphid); Eriosoma lanigerum Hausmann (woolly appleaphid); Brevicoryne brassicae Linnaeus (cabbage aphid); Hyalopteruspruni Geoffroy (mealy plum aphid); Lipaphis erysimi Kaltenbach (turnipaphid); Metopolophium dirrhodum Walker (cereal aphid); Macrosiphumeuphorbiae Thomas (potato aphid); Myzus persicae Sulzer (peach-potatoaphid, green peach aphid); Nasonovia ribisnigri Mosley (lettuce aphid);Pemphigus spp. (root aphids and gall aphids); Rhopalosiphum maidis Fitch(corn leaf aphid); R. padi Linnaeus (bird cherry-oat aphid); Schizaphisgraminum Rondani (greenbug); Sipha flava Forbes (yellow sugarcaneaphid); Sitobion avenae Fabricius (English grain aphid); Therioaphismaculata Buckton (spotted alfalfa aphid); Toxoptera aurantii Boyer deFonscolombe (black citrus aphid) and T. citricida Kirkaldy (brown citrusaphid); Adelges spp. (adelgids); Phylloxera devastatrix Pergande (pecanphylloxera); Bemisia tabaci Gennadius (tobacco whitefly, sweetpotatowhitefly); B. argentifolii Bellows & Perring (silverleaf whitefly);Dialeurodes citri Ashmead (citrus whitefly); Trialeurodes abutiloneus(bandedwinged whitefly) and T. vaporariorum Westwood (greenhousewhitefly); Empoasca fabae Harris (potato leafhopper); Laodelphaxstriatellus Fallen (smaller brown planthopper); Macrolestesquadrilineatus Forbes (aster leafhopper); Nephotettix cinticeps Uhler(green leafhopper); N. nigropictus Stal (rice leafhopper); Nilaparvatalugens Stal (brown planthopper); Peregrinus maidis Ashmead (cornplanthopper); Sogatella furcifera Horvath (white-backed planthopper);Sogatodes orizicola Muir (rice delphacid); Typhlocyba pomaria McAtee(white apple leafhopper); Erythroneoura spp. (grape leafhoppers);Magicicada septendecim Linnaeus (periodical cicada); Icerya purchasiMaskell (cottony cushion scale); Quadraspidiotus perniciosus Comstock(San Jose scale); Planococcus citri Risso (citrus mealybug);Pseudococcus spp. (other mealybug complex); Cacopsylla pyricola Foerster(pear psylla); Trioza diospyri Ashmead (persimmon psylla).

Agronomically important species of interest from the order Hemipterainclude, but are not limited to: Acrosternum hilare Say (green stinkbug); Anasa tristis De Geer (squash bug); Blissus leucopterusleucopterus Say (chinch bug); Corythuca gossypii Fabricius (cotton lacebug); Cyrtopeltis modesta Distant (tomato bug); Dysdercus suturellusHerrich-Schaffer (cotton stainer); Euschistus servus Say (brown stinkbug); E. variolarius Palisot de Beauvois (one-spotted stink bug);Graptostethus spp. (complex of seed bugs); Leptoglossus corculus Say(leaf-footed pine seed bug); Lygus lineolaris Palisot de Beauvois(tarnished plant bug); L. Hesperus Knight (Western tarnished plant bug);L. pratensis Linnaeus (common meadow bug); L. rugulipennis Poppius(European tarnished plant bug); Lygocoris pabulinus Linnaeus (commongreen capsid); Nezara viridula Linnaeus (southern green stink bug);Oebalus pugnax Fabricius (rice stink bug); Oncopeltus fasciatus Dallas(large milkweed bug); Pseudatomoscelis seriatus Reuter (cottonfleahopper).

Furthermore, embodiments may be effective against Hemiptera such,Calocoris norvegicus Gmelin (strawberry bug); Orthops campestrisLinnaeus; Plesiocoris rugicollis Fallen (apple capsid); Cyrtopeltismodestus Distant (tomato bug); Cyrtopeltis notatus Distant (suckfly);Spanagonicus albofasciatus Reuter (whitemarked fleahopper); Diaphnocorischlorionis Say (honeylocust plant bug); Labopidicola allii Knight (onionplant bug); Pseudatomoscelis seriatus Reuter (cotton fleahopper);Adelphocoris rapidus Say (rapid plant bug); Poecilocapsus lineatusFabricius (four-lined plant bug); Nysius ericae Schilling (false chinchbug); Nysius raphanus Howard (false chinch bug); Nezara viridulaLinnaeus (Southern green stink bug); Eurygaster spp.; Coreidae spp.;Pyrrhocoridae spp.; Tinidae spp.; Blostomatidae spp.; Reduviidae spp.and Cimicidae spp.

Also included are adults and larvae of the order Acari (mites) such asAceria tosichella Keifer (wheat curl mite); Petrobia latens Müller(brown wheat mite); spider mites and red mites in the familyTetranychidae, Panonychus ulmi Koch (European red mite); Tetranychusurticae Koch (two spotted spider mite); (T. mcdanieli McGregor (McDanielmite); T. cinnabarinus Boisduval (carmine spider mite); T. turkestaniUgarov & Nikolski (strawberry spider mite); flat mites in the familyTenuipalpidae, Brevipalpus lewisi McGregor (citrus flat mite); rust andbud mites in the family Eriophyidae and other foliar feeding mites andmites important in human and animal health, i.e., dust mites in thefamily Epidermoptidae, follicle mites in the family Demodicidae, grainmites in the family Glycyphagidae, ticks in the order Ixodidae. Ixodesscapularis Say (deer tick); I. holocyclus Neumann (Australian paralysistick); Dermacentor variabilis Say (American dog tick); Amblyommaamericanum Linnaeus (lone star tick) and scab and itch mites in thefamilies Psoroptidae, Pyemotidae and Sarcoptidae.

Insect pests of the order Thysanura are of interest, such as Lepismasaccharina Linnaeus (silverfish); Thermobia domestica Packard(firebrat).

Additional arthropod pests covered include: spiders in the order Araneaesuch as Loxosceles reclusa Gertsch and Mulaik (brown recluse spider) andthe Latrodectus mactans Fabricius (black widow spider) and centipedes inthe order Scutigeromorpha such as Scutigera coleoptrata Linnaeus (housecentipede).

Insect pest of interest include the superfamily of stink bugs and otherrelated insects including but not limited to species belonging to thefamily Pentatomidae (Nezara viridula, Halyomorpha halys, Piezodorusguildini, Euschistus servus, Acrosternum hilare, Euschistus heros,Euschistus tristigmus, Acrosternum hilare, Dichelops furcatus, Dichelopsmelacanthus, and Bagrada hilaris (Bagrada Bug)), the family Plataspidae(Megacopta cribraria—Bean plataspid) and the family Cydnidae(Scaptocoris castanea—Root stink bug) and Lepidoptera species includingbut not limited to: diamond-back moth, e.g., Helicoverpa zea Boddie;soybean looper, e.g., Pseudoplusia includens Walker and velvet beancaterpillar e.g., Anticarsia gemmatalis Hübner.

Methods for measuring pesticidal activity are well known in the art.See, for example, Czapla and Lang, (1990) J. Econ. Entomol.83:2480-2485; Andrews, et al., (1988) Biochem. J. 252:199-206; Marrone,et al., (1985) J. of Economic Entomology 78:290-293 and U.S. Pat. No.5,743,477, all of which are herein incorporated by reference in theirentirety. Generally, the protein is mixed and used in feeding assays.See, for example Marrone, et al., (1985) J. of Economic Entomology78:290-293. Such assays can include contacting plants with one or morepests and determining the plant's ability to survive and/or cause thedeath of the pests.

Nematodes include parasitic nematodes such as root-knot, cyst and lesionnematodes, including Heterodera spp., Meloidogyne spp. and Globoderaspp.; particularly members of the cyst nematodes, including, but notlimited to, Heterodera glycines (soybean cyst nematode); Heteroderaschachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode)and Globodera rostochiensis and Globodera pailida (potato cystnematodes). Lesion nematodes include Pratylenchus spp.

Seed Treatment

To protect and to enhance yield production and trait technologies, seedtreatment options can provide additional crop plan flexibility and costeffective control against insects, weeds and diseases. Seed material canbe treated with one or more of the insecticidal proteins or polypeptidesdisclosed herein. For e.g., such seed treatments can be applied on seedsthat contain a transgenic trait including transgenic corn, soy,brassica, cotton or rice. Combinations of one or more of theinsecticidal proteins or polypeptides disclosed herein and otherconventional seed treatments are contemplated. Seed material can betreated, typically surface treated, with a composition comprisingcombinations of chemical or biological herbicides, herbicide safeners,insecticides, fungicides, germination inhibitors and enhancers,nutrients, plant growth regulators and activators, bactericides,nematocides, avicides and/or molluscicides. These compounds aretypically formulated together with further carriers, surfactants orapplication-promoting adjuvants customarily employed in the art offormulation. The coatings may be applied by impregnating propagationmaterial with a liquid formulation or by coating with a combined wet ordry formulation. Examples of the various types of compounds that may beused as seed treatments are provided in The Pesticide Manual: A WorldCompendium, C. D. S. Tomlin Ed., Published by the British CropProduction Council, which is hereby incorporated by reference.

Some seed treatments that may be used on crop seed include, but are notlimited to, one or more of abscisic acid, acibenzolar-S-methyl,avermectin, amitrol, azaconazole, azospirillum, azadirachtin,azoxystrobin, Bacillus spp. (including one or more of cereus, firmus,megaterium, pumilis, sphaericus, subtilis and/or thuringiensis species),bradyrhizobium spp. (including one or more of betae, canariense,elkanii, iriomotense, japonicum, liaonigense, pachyrhizi and/oryuanmingense), captan, carboxin, chitosan, clothianidin, copper,cyazypyr, difenoconazole, etidiazole, fipronil, fludioxonil,fluoxastrobin, fluquinconazole, flurazole, fluxofenim, harpin protein,imazalil, imidacloprid, ipconazole, isoflavenoids,lipo-chitooligosaccharide, mancozeb, manganese, maneb, mefenoxam,metalaxyl, metconazole, myclobutanil, PCNB, penflufen, penicillium,penthiopyrad, permethrine, picoxystrobin, prothioconazole,pyraclostrobin, rynaxypyr, S-metolachlor, saponin, sedaxane, TCMTB,tebuconazole, thiabendazole, thiamethoxam, thiocarb, thiram,tolclofos-methyl, triadimenol, trichoderma, trifloxystrobin,triticonazole and/or zinc. PCNB seed coat refers to EPA RegistrationNumber 00293500419, containing quintozen and terrazole. TCMTB refers to2-(thiocyanomethylthio) benzothiazole.

Seed varieties and seeds with specific transgenic traits may be testedto determine which seed treatment options and application rates maycomplement such varieties and transgenic traits in order to enhanceyield. For example, a variety with good yield potential but head smutsusceptibility may benefit from the use of a seed treatment thatprovides protection against head smut, a variety with good yieldpotential but cyst nematode susceptibility may benefit from the use of aseed treatment that provides protection against cyst nematode, and soon. Likewise, a variety encompassing a transgenic trait conferringinsect resistance may benefit from the second mode of action conferredby the seed treatment, a variety encompassing a transgenic traitconferring herbicide resistance may benefit from a seed treatment with asafener that enhances the plants resistance to that herbicide, etc.Further, the good root establishment and early emergence that resultsfrom the proper use of a seed treatment may result in more efficientnitrogen use, a better ability to withstand drought and an overallincrease in yield potential of a variety or varieties containing acertain trait when combined with a seed treatment.

Methods for Killing an Insect Pest and Controlling an Insect Population

In some embodiments methods are provided for killing an insect pest,comprising contacting the insect pest, either simultaneously orsequentially, with an insecticidally-effective amount of a recombinantPtIP-50 polypeptide and PtIP-65 polypeptide. In some embodiments methodsare provided for killing an insect pest, comprising contacting theinsect pest with an insecticidally-effective amount of a recombinantpesticidal protein of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ IDNO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 orSEQ ID NO: 70 and/or PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO:42 or a variant thereof.

In some embodiments methods are provided for controlling an insect pestpopulation, comprising contacting the insect pest population, eithersimultaneously or sequentially, with an insecticidally-effective amountof a recombinant PtIP-50 polypeptide and PtIP-65 polypeptide. In someembodiments methods are provided for controlling an insect pestpopulation, comprising contacting the insect pest population with aninsecticidally-effective amount of a recombinant PtIP-50 polypeptide ofSEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO:47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ IDNO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61,SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO:66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ ID NO: 70 and/orPtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29,SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO:34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ IDNO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42 or a variantthereof. As used herein, “controlling a pest population” or “controls apest” refers to any effect on a pest that results in limiting the damagethat the pest causes. Controlling a pest includes, but is not limitedto, killing the pest, inhibiting development of the pest, alteringfertility or growth of the pest in such a manner that the pest providesless damage to the plant, decreasing the number of offspring produced,producing less fit pests, producing pests more susceptible to predatorattack or deterring the pests from eating the plant.

In some embodiments methods are provided for controlling an insect pestpopulation resistant to a pesticidal protein, comprising contacting theinsect pest population, either simultaneously or sequentially, with aninsecticidally-effective amount of a recombinant PtIP-50 polypeptide anda recombinant PtIP-65 polypeptide. In some embodiments methods areprovided for controlling an insect pest population resistant to apesticidal protein, comprising contacting the insect pest populationwith an insecticidally-effective amount of a recombinant PtIP-50polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO:46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ IDNO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO:65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ IDNO: 70 and/or the PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ IDNO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42or a variant thereof.

In some embodiments methods are provided for protecting a plant from aninsect pest, comprising expressing in the plant or cell thereof at leastone recombinant polynucleotide encoding a PtIP-50 polypeptide and arecombinant PtIP-65 polypeptide. In some embodiments methods areprovided for protecting a plant from an insect pest, comprisingexpressing in the plant or cell thereof a recombinant polynucleotideencoding PtIP-50 polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ IDNO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59,SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ IDNO: 69 or SEQ ID NO: 70 and/or PtIP-65 polypeptide of SEQ ID NO: 22, SEQID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27,SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO:32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ IDNO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 orSEQ ID NO: 42 or variants thereof.

In some embodiments methods are provided for protecting a plant from aninsect pest, comprising expressing in the plant or cell thereof at leastone recombinant polynucleotide encoding a PtIP-50 polypeptide and arecombinant PtIP-65 polypeptide. In some embodiments methods areprovided for protecting a plant from an insect pest, comprisingexpressing in the plant or cell thereof a recombinant polynucleotideencoding a PtIP-50 polypeptide of any one of SEQ ID NO: 224-326.

Insect Resistance Management (IRM) Strategies

Expression of B. thuringiensis δ-endotoxins in transgenic corn plantshas proven to be an effective means of controlling agriculturallyimportant insect pests (Perlak, et al., 1990; 1993). However, insectshave evolved that are resistant to B. thuringiensis δ-endotoxinsexpressed in transgenic plants. Such resistance, should it becomewidespread, would clearly limit the commercial value of germplasmcontaining genes encoding such B. thuringiensis 5-endotoxins.

One way to increasing the effectiveness of the transgenic insecticidesagainst target pests and contemporaneously reducing the development ofinsecticide-resistant pests is to use provide non-transgenic (i.e.,non-insecticidal protein) refuges (a section of non-insecticidalcrops/corn) for use with transgenic crops producing a singleinsecticidal protein active against target pests. The United StatesEnvironmental Protection Agency(epa.gov/oppbppdl/biopesticides/pips/bt_corn_refuge_2006.htm, which canbe accessed using the www prefix) publishes the requirements for usewith transgenic crops producing a single Bt protein active againsttarget pests. In addition, the National Corn Growers Association, ontheir website:(ncga.com/insect-resistance-management-fact-sheet-bt-corn, which can beaccessed using the www prefix) also provides similar guidance regardingrefuge requirements. Due to losses to insects within the refuge area,larger refuges may reduce overall yield.

Another way of increasing the effectiveness of the transgenicinsecticides against target pests and contemporaneously reducing thedevelopment of insecticide-resistant pests would be to have a repositoryof insecticidal genes that are effective against groups of insect pestsand which manifest their effects through different modes of action.

Expression in a plant of two or more insecticidal compositions toxic tothe same insect species, each insecticide being expressed at efficaciouslevels would be another way to achieve control of the development ofresistance. This is based on the principle that evolution of resistanceagainst two separate modes of action is far more unlikely than only one.Roush, for example, outlines two-toxin strategies, also called“pyramiding” or “stacking,” for management of insecticidal transgeniccrops. (The Royal Society. Phil. Trans. R. Soc. Lond. B. (1998)353:1777-1786). Stacking or pyramiding of two different proteins eacheffective against the target pests and with little or nocross-resistance can allow for use of a smaller refuge. The USEnvironmental Protection Agency requires significantly less (generally5%) structured refuge of non-Bt corn be planted than for single traitproducts (generally 20%). There are various ways of providing the IRMeffects of a refuge, including various geometric planting patterns inthe fields and in-bag seed mixtures, as discussed further by Roush.

In some embodiments the PtIP-50 polypeptide and PtIP-65 polypeptide ofthe disclosure are useful as an insect resistance management strategy incombination (i.e., pyramided) with other pesticidal proteins include butare not limited to Bt toxins, Xenorhabdus sp. or Photorhabdus sp.insecticidal proteins, and the like.

Provided are methods of controlling Lepidoptera and/or Coleoptera insectinfestation(s) in a transgenic plant that promote insect resistancemanagement, comprising expressing in the plant at least two differentinsecticidal proteins having different modes of action.

In some embodiments the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management the at least one of the insecticidal proteinscomprise a PtIP-50 polypeptide and PtIP-65 polypeptide insecticidal toinsects in the order Lepidoptera and/or Coleoptera.

In some embodiments the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprise expressing at least one of the PtIP-50polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO:46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ IDNO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO:65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 or SEQ IDNO: 70 and/or a PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO:33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ IDNO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 42 orvariants thereof, insecticidal to insects in the order Lepidopteraand/or Coleoptera.

In some embodiments the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprise expressing in the transgenic plant aPtIP-50 polypeptide and PtIP-65 polypeptide and a Cry proteininsecticidal to insects in the order Lepidoptera and/or Coleopterahaving different modes of action.

In some embodiments the methods of controlling Lepidoptera and/orColeoptera insect infestation in a transgenic plant and promoting insectresistance management comprise expressing in the transgenic plant aPtIP-50 polypeptide of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ IDNO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69 orSEQ ID NO: 70 and/or a PtIP-65 polypeptide of SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37,SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO:42 or variants thereof and a Cry protein insecticidal to insects in theorder Lepidoptera and/or Coleoptera having different modes of action.

Also provided are methods of reducing likelihood of emergence ofLepidoptera and/or Coleoptera insect resistance to transgenic plantsexpressing in the plants insecticidal proteins to control the insectspecies, comprising expression of a PtIP-50 polypeptide and a PtIP-65polypeptide insecticidal to the insect species in combination with asecond insecticidal protein to the insect species having different modesof action.

Also provided are means for effective Lepidoptera and/or Coleopterainsect resistance management of transgenic plants, comprisingco-expressing at high levels in the plants two or more insecticidalproteins toxic to Lepidoptera and/or Coleoptera insects but eachexhibiting a different mode of effectuating its killing activity,wherein the two or more insecticidal proteins comprise a PtIP-50polypeptide and PtIP-65 polypeptide and a Cry protein. Also provided aremeans for effective Lepidoptera and/or Coleoptera insect resistancemanagement of transgenic plants, comprising co-expressing at high levelsin the plants two or more insecticidal proteins toxic to Lepidopteraand/or Coleoptera insects but each exhibiting a different mode ofeffectuating its killing activity, wherein the two or more insecticidalproteins comprise a PtIP-50 polypeptide of SEQ ID NO: 43, SEQ ID NO: 44,SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO:49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ IDNO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63,SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO:68, SEQ ID NO: 69 or SEQ ID NO: 70 and/or PtIP-65 polypeptide of SEQ IDNO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31,SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO:36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ IDNO: 41 or SEQ ID NO: 42 or variants thereof and a Cry protein.

In addition, methods are provided for obtaining regulatory approval forplanting or commercialization of plants expressing proteins insecticidalto insects in the order Lepidoptera and/or Coleoptera, comprising thestep of referring to, submitting or relying on insect assay binding datashowing that the PtIP-50 polypeptide and the PtIP-65 polypeptide doesnot compete with binding sites for Cry proteins in such insects. Inaddition, methods are provided for obtaining regulatory approval forplanting or commercialization of plants expressing proteins insecticidalto insects in the order Lepidoptera and/or Coleoptera, comprising thestep of referring to, submitting or relying on insect assay binding datashowing that the PtIP-50 polypeptide of SEQ ID NO: 43, SEQ ID NO: 44,SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO:49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ IDNO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63,SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO:68, SEQ ID NO: 69 or SEQ ID NO: 70 and/or the PtIP-65 polypeptide of SEQID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26,SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ IDNO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQID NO: 41 or SEQ ID NO: 42 or variant thereof does not compete withbinding sites for Cry proteins in such insects.

Methods for Increasing Plant Yield

Methods for increasing plant yield are provided. The methods compriseproviding a plant or plant cell expressing a polynucleotide encoding thepesticidal polypeptide sequence disclosed herein and growing the plantor a seed thereof in a field infested with a pest against which thepolypeptide has pesticidal activity. In some embodiments, thepolypeptide has pesticidal activity against a Lepidopteran, Coleopteran,Dipteran, Hemipteran or nematode pest, and the field is infested with aLepidopteran, Hemipteran, Coleopteran, Dipteran or nematode pest.

As defined herein, the “yield” of the plant refers to the quality and/orquantity of biomass produced by the plant. “Biomass” as used hereinrefers to any measured plant product. An increase in biomass productionis any improvement in the yield of the measured plant product.Increasing plant yield has several commercial applications. For example,increasing plant leaf biomass may increase the yield of leafy vegetablesfor human or animal consumption. Additionally, increasing leaf biomasscan be used to increase production of plant-derived pharmaceutical orindustrial products. An increase in yield can comprise any statisticallysignificant increase including, but not limited to, at least a 1%increase, at least a 3% increase, at least a 5% increase, at least a 10%increase, at least a 20% increase, at least a 30%, at least a 50%, atleast a 70%, at least a 100% or a greater increase in yield compared toa plant not expressing the pesticidal sequence.

In specific methods, plant yield is increased as a result of improvedpest resistance of a plant expressing a PtIP-50 polypeptide and aPtIP-65 polypeptide disclosed herein. Expression of the PtIP-50polypeptide and the PtIP-65 polypeptide results in a reduced ability ofa pest to infest or feed on the plant, thus improving plant yield.

Methods of Processing

Further provided are methods of processing a plant, plant part or seedto obtain a food or feed product from a plant, plant part or seedcomprising a PtIP-50 polypeptide and a PtIP-65 polypeptide. The plants,plant parts or seeds provided herein, can be processed to yield oil,protein products and/or by-products that are derivatives obtained byprocessing that have commercial value. Non-limiting examples includetransgenic seeds comprising a nucleic acid molecule encoding a PtIP-50polypeptide and a PtIP-65 polypeptide which can be processed to yieldsoy oil, soy products and/or soy by-products.

“Processing” refers to any physical and chemical methods used to obtainany soy product and includes, but is not limited to, heat conditioning,flaking and grinding, extrusion, solvent extraction or aqueous soakingand extraction of whole or partial seeds

The following examples are offered by way of illustration and not by wayof limitation.

EXPERIMENTALS Example 1—Identification of an Insecticidal Protein Activefrom the Fern, Asplenium australasicum, (PS-8566)

The insecticidal proteins PtIP-50Aa and PtIP-65Aa were identified byprotein purification, mass spectroscopy (MS) and PCR cloning fromAsplenium australasicum, PS-8566 as follows.

Asplenium australasicum was collected by a collaborator and assignedidentification number PS-8566. A voucher specimen of Aspleniumaustralasicum was sent to the Delaware State University Herbarium wherethe identity was verified. PS-8566 was collected, flash frozen in liquidN₂ and stored at −80° C. After storage it was ground to a fine powder atliquid N₂ temperatures with a Geno Ball Mill (SPEX, Metuchen, N.J.). Toextract protein, 20 ml of 50 mM Tris buffer, pH 8.0, 150 mM KCl, 2.5 mMEDTA, 1.5% polyvinylpolypyrrolidone (PVPP) and protease inhibitorcocktail (Roche Diagnostics, Germany) was added to every 5 g freshweight of tissue. The homogenate was centrifuged to remove cell debris,filtered through 0.22 um filters and desalted using 10 ml Zeba SpinDesalting columns (Thermo Scientific, IL.)

Bioassays against the three pest species, Soybean Looper (SBL)(Pseudoplusia includens), Corn Earworm (CEW) (Helicoverpa zea) andEuropean Corn Borer (ECB) (Ostrinia nubialis) were conducted using thedesalted protein extract overlaid onto an agar-based Lepidoptera diet(Southland Products Inc., Lake Village, Ark.) in a 96-well plate format.

Six replicates were used per sample. Samples were allowed to dry on topof the diet and two to five neonate insects were placed into each wellof the treated plate. After four days of incubation at 27° C. larvaewere scored for mortality or severity of stunting. The scores wererecorded numerically as dead (3), severely stunted (2) (little or nogrowth but alive and equivalent to a 1^(st) instar larvae), stunted (1)(growth to second instar but not equivalent to controls), or normal (0).Subjecting the sample to proteinase K and heat treatments resulted inloss of activity indicating that the sample was proteinaceous in nature.Bioassay results are shown in Table 7.

TABLE 7 Activity of A. australasicum protein extract against lepidopteralarvae Ave. Ave. Score after Score Proteinase K/Heat Neonate SoybeanLooper 3 0 Corn Earworm 2 0 European Corn Borer 1.5 0

For protein purification, PS-8566 was ground to a fine powder at liquidN₂ temperatures with a Geno Ball Mill (SPEX, Metuchen, N.J.). Proteinwas extracted in 50 mM Tris buffer, pH 8.0, 150 mM KCl, 2.5 mM EDTA,1.5% PVPP and protease inhibitor cocktail (Roche Diagnostics, Germany).The extracted material was centrifuged to remove cell debris, filteredthrough 0.22 um filters and desalted using a HiPrep 26/10 desaltingcolumn (GE, Piscataway, N.J.). The desalted fractions were assayedagainst CEW and SBL as described above. The active protein fraction poolwas loaded onto a 5 ml HiTrap Q column (GE, Piscataway, N.J.) and elutedwith a linear (30 CV) gradient from 0 M to 0.6 M NaCl in 50 mM Tris, pH8.0, The active fractions were combined, concentrated by Amiconmolecular weight cutoff filtration (Millipore, Billerica, Mass.), andloaded onto a HiLoad™ Superdex™ 200, 16/60 Size exclusion column (GE,Piscataway, N.J.) and eluted with 50 mM Tris buffer, pH 8.0, 150 mMNaCl. Fractions active against SBL and CEW were combined, concentratedand then further diluted with 50 mM Tris buffer, pH 8 to reduce the saltconcentration and loaded onto a Mono Q® 5/50 column (GE, PiscatawayN.J.). Fractions eluted from the Mono Q® 5/50 column with a 0 to 0.7MNaCl gradient over 200 CV were assayed against SBL and CEW. Based onLDS-PAGE the active fraction contained two protein bands atapproximately 89 kDa and 35 kDa. The two bands were designated withinternal names to be used hereafter. The protein representing the 89 kDaband was named PtIP-50Aa (SEQ ID NO: 71) and the protein representingthe 35 kDa band was named PtIP-65Aa (SEQ ID NO: 22).

Protein sequencing and identification was performed by MS analysis afterprotein digestion with trypsin. Proteins for MS identification wereobtained after running the sample on an LDS-PAGE gel stained withBrilliant Blue G-250 Stain. The two bands of interest were excised fromthe gel, de-stained, reduced with dithiothreitol and then alkylated withiodoacetamide. Following overnight digestion with trypsin, the sampleswere submitted for LCMS analysis. Liquid chromatography-tandem massspectrometry (LC-MSMS) analysis for tryptically-digested peptides wasconducted using electrospray on a QToF Premiere™ mass spectrometer(Waters®, Milford, Mass.) coupled with a NanoAcquity™ nano-LC system(Waters®, Milford, Mass.) with a gradient from 2% acetonitrile, 0.1%formic acid to 60% acetonitrile, 0.1% formic acid.

The resulting LCMS data were analyzed using Protein Lynx Global Server(Waters®, Milford, Mass.) to generate DeNovo sequence data. The aminoacid sequences were BLAST (Basic Local Alignment Search Tool; Altschul,et al., (1993) J. Mol. Biol. 215:403-410; see alsoncbi.nlm.nih.gov/BLAST/, which can be accessed using the www prefix)searched against public and DUPONT-PIONEER internal databases thatincluded plant protein sequences. Amino acid sequences were aligned withproteins in a proprietary DUPONT-PIONEER plant protein database.

Example 2—Transcriptomic Sequencing of PtIP-50 and PtIP-65

A transcriptome for Asplenium australasicum, PS-8566 was prepared asfollows. Total RNAs were isolated from frozen tissues by use of theQiagen® RNeasy® kit for total RNA isolation. Sequencing libraries fromthe resulting total RNAs were prepared using the TruSeq™ mRNA-Seq kitand protocol from Illumina®, Inc. (San Diego, Calif.). Briefly, mRNAswere isolated via attachment to oligo(dT) beads, fragmented to a meansize of 180 nt, reverse transcribed into cDNA by random hexamer prime,end repaired, 3′ A-tailed, and ligated with Illumina® indexed TruSeq™adapters. Ligated cDNA fragments were PCR amplified using Illumina®TruSeq™ primers and purified PCR products were checked for quality andquantity on the Agilent Bioanalyzer® DNA 7500 chip. Post quality andquantity assessment, 100 ng of the transcript library was normalized bytreatment with Duplex Specific Nuclease (DSN) (Evrogen®, Moscow,Russia). Normalization was accomplished by addition of 200 mM Hepesbuffer, followed by heat denaturation and five hour anneal at 68° C.Annealed library was treated with 2 ul of DSN enzyme for 25 minutes,purified by Qiagen® MinElute® columns according to manufacturerprotocols, and amplified twelve cycles using Illumina® adapter specificprimers. Final products were purified with Ampure® XP beads (BeckmanGenomics, Danvers, Mass.) and checked for quality and quantity on theAgilent Bioanalyzer® DNA 7500 chip.

Normalized transcript libraries were sequenced according to manufacturerprotocols on the Illumina® Genome Analyzer IIx. Each library washybridized to two flowcell lanes and amplified, blocked, linearized andprimer hybridized using the Illumina clonal cluster generation processon cBot®. Sequencing was completed on the Genome Analyzer IIx,generating sixty million 75 bp paired end reads per normalized library.

Peptide sequences identified for PtIP-50Aa (SEQ ID NO: 71) byLC-MS/MS/MS sequencing (described in Example 1) were searched againstthe protein sequences predicted by open reading frames (ORFs) from theinternal transcriptome for PS-8566 assemblies. The peptides gave aperfect match to a transcript corresponding to PtIP-50Aa (SEQ ID NO:71). Amino acid sequence stretches identified for PtIP-65Aa (SEQ ID NO:22) by LC-MS/MS/MS sequencing were searched against the proteinspredicted by open reading frames (ORFs) of the contig assemblies. Thepeptides gave perfect matches to another transcript corresponding toPtIP-65Aa (SEQ ID NO: 22).

The coding sequences were used to design the following primers to clonethe PtIP-50Aa coding sequence: CGAATTCGAGCACATATGGCCGCC (95KD N-T Ndelfor) (SEQ ID NO: 202) and GATTCCGGATCCTTATACGGCGACTCTGG (95KD C-T BamHIREV) (SEQ ID NO: 203). This clone was produced by polymerase chainreaction using the HF Advantage® PCR kit (Clontech™, 1290 Terra BellaAve. Mountain View, Calif. 94043) and the total RNA from Aspleniumaustralasicum as a template. The cloned sequence was confirmed bysequencing.

The coding sequences were used to design the following primers to clonethe PtIP-65Aa coding sequence: GGATACACCTCATATGGCGTTGGTGATCGG (36KD N-TNdel for) (SEQ ID NO: 204) and GTGCGGGATCCCTACTGAGTGGTGATGTCAATCGCG(36KD C-T BamHI REV) (SEQ ID NO: 205). This clone was produced bypolymerase chain reaction using the HF Advantage® PCR kit (Clontech™,1290 Terra Bella Ave. Mountain View, Calif. 94043) and the total RNAfrom Asplenium australasicum as a template. The cloned sequence wasconfirmed by sequencing.

Based on the DNA and protein sequencing, the PtIP-50Aa polynucleotidesequence is provided as SEQ ID NO: 43 and the polypeptide sequence asSEQ ID NO: 71. The PtIP-65Aa polynucleotide sequence is provided as SEQID NO: 1 and the polypeptide sequence as SEQ ID NO: 22.

Example 3:—Identification of PtIP-50 and PtIP-65 Homologs

Gene identities may be determined by conducting BLAST (Basic LocalAlignment Search Tool; Altschul, et al., (1993) J. Mol. Biol.215:403-410; see also ncbi.nlm.nih.gov/BLAST/, which can be accessedusing the www prefix) searches under default parameters for similarityto sequences. The polynucleotide sequences for PtIP-50Aa (SEQ ID NO: 43)and PtIP-65Aa (SEQ ID NO: 1) were analyzed. Gene identities conducted byBLAST in a DUPONT PIONEER internal transcriptomes database of ferns andother primitive plants identified multiple homologs for both PtIP-50Aa(SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) within a range ofhomologies. Table 8 shows the Plant Sample ID #, fern or other primitiveplant species, internal clone designation and the respective sequenceidentifiers for the PtIP-65 polypeptide homologs and polynucleotidesencoding the PtIP-50 homologs. Table 9 shows the Plant Sample ID #, fernor other primitive plant species, internal clone designation and therespective sequence identifiers for the PtIP-65 polypeptide homologs andpolynucleotides encoding the PtIP-50 homologs.

TABLE 8 Plant Sample ID Species name n.a. seq a.a. seq PS-8566 Aspleniumaustralasicum PtIP-65Aa SEQ ID NO: 1 SEQ ID NO: 22 PS-9140 Asplenium ×kenzoi PtIP-65Hc SEQ ID NO: 2 SEQ ID NO: 23 Sa. Kurata PS-8568Polypodium musifolium PtIP-65Ba SEQ ID NO: 3 SEQ ID NO: 24 PtIP-65Ha SEQID NO: 4 SEQ ID NO: 25 PS-9319 Polypodium punctatum PtIP-65Bb SEQ ID NO:5 SEQ ID NO: 26 ‘Serratum’ PtIP-65Gc SEQ ID NO: 6 SEQ ID NO: 27PtIP-65Hd SEQ ID NO: 7 SEQ ID NO: 28 PS-7140 Adiantum pedatum L.PtIP-65Ca SEQ ID NO: 8 SEQ ID NO: 29 PS-9135 Platycerium bifurcatumPtIP-65Ga SEQ ID NO: 9 SEQ ID NO: 30 PtIP-65Hb SEQ ID NO: 10 SEQ ID NO:31 PS-8837 Nephrolepis falcata PtIP-65He SEQ ID NO: 11 SEQ ID NO: 32PtIP-65Hf SEQ ID NO: 12 SEQ ID NO: 33 PS-9145 Ophioglossum pendulumPtIP-65Gb SEQ ID NO: 13 SEQ ID NO: 34 PS-9427 Colysis wrightii PtIP-65GdSEQ ID NO: 14 SEQ ID NO: 35 ‘Monstifera’ PS-7897 Colysis wrightiiPtIP-65Ge SEQ ID NO: 15 SEQ ID NO: 36 PtIP-65Hk SEQ ID NO: 16 SEQ ID NO:37 PS-8780 Selaginella kraussiana PtIP-65Hh SEQ ID NO: 17 SEQ ID NO: 38‘Variegata’ PtIP-65Hj SEQ ID NO: 18 SEQ ID NO: 39 PS-7896 Selaginellavictoriae PtIP-65Hg SEQ ID NO: 19 SEQ ID NO: 40 PS-8582 Lycopodiumphlegmaria PtIP-65Fa SEQ ID NO: 20 SEQ ID NO: 41 PS-9141 Huperziasalvinioides PtIP-65Fb SEQ ID NO: 21 SEQ ID NO: 42

TABLE 9 Plant Sample ID Species name n.a. seq a.a. seq PS-8566 Aspleniumaustralasicum PtIP-50Aa SEQ ID NO: 43 SEQ ID NO: 71 PtIP-50Fb SEQ ID NO:44 SEQ ID NO: 72 PS-8564 Asplenium nidus PtIP-50Fo SEQ ID NO: 45 SEQ IDNO: 73 PS-9140 Asplenium × kenzoi PtIP-50Fs SEQ ID NO: 46 SEQ ID NO: 74Sa. Kurata PtIP-50Fr SEQ ID NO: 47 SEQ ID NO: 75 PtIP-50Gc SEQ ID NO: 48SEQ ID NO: 76 PS-8568 Polypodium musifolium PtIP-50Ba SEQ ID NO: 49 SEQID NO: 77 PS-9319 Polypodium punctatum PtIP-50Bc SEQ ID NO: 50 SEQ IDNO: 78 ‘Serratum’ PtIP-50Fe SEQ ID NO: 51 SEQ ID NO: 79 PtIP-50Gb SEQ IDNO: 52 SEQ ID NO: 78 PtIP-50Fh SEQ ID NO: 53 SEQ ID NO: 81 PtIP-50Fg SEQID NO: 54 SEQ ID NO: 82 PtIP-50Ff SEQ ID NO: 55 SEQ ID NO: 83 PS-7140Adiantum pedatum L. PtIP-50Bb SEQ ID NO: 56 SEQ ID NO: 84 PtIP-50Ft SEQID NO: 57 SEQ ID NO: 85 PtIP-50Fa SEQ ID NO: 58 SEQ ID NO: 86 PS-9135Platycerium bifurcatum PtIP-50Fd SEQ ID NO: 59 SEQ ID NO: 87 PtIP-50GaSEQ ID NO: 60 SEQ ID NO: 88 PS-8837 Nephrolepis falcata PtIP-50Fk SEQ IDNO: 61 SEQ ID NO: 89 PtIP-50Fi SEQ ID NO: 62 SEQ ID NO: 90 PtIP-50Fj SEQID NO: 63 SEQ ID NO: 91 PtIP-50Gd SEQ ID NO: 64 SEQ ID NO: 92 PS-9427Colysis wrightii PtIP-50Fn SEQ ID NO: 65 SEQ ID NO: 93 ‘Monstifera’PS-7897 Colysis wrightii PtIP-50Fl SEQ ID NO: 66 SEQ ID NO: 94 PtIP-50FqSEQ ID NO: 67 SEQ ID NO: 95 PS-9321 Bolbitis cladorrhizans PtIP-50Bd SEQID NO: 68 SEQ ID NO: 96 PtIP-50Fp SEQ ID NO: 69 SEQ ID NO: 97 PS-9537Blechnum brasiliense PtIP-50Fm SEQ ID NO: 70 SEQ ID NO: 98 ‘Crispum’

TABLE 10 PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP-PtIP- PtIP- PtIP- PtIP- 50Ba 50Bb 50Bc 50Bd 50Fa 50Fb 50Fd 50Fe 50Ff50Fg 50Fh 50Fi 50Fj 50Fk PtIP-50Aa 86.3 84.8 83.0 88.4 46.3 48.4 46.948.9 51.3 46.1 47.7 48.4 50.9 46.1 PtIP-50Ba — 79.3 87.5 81.3 45.7 47.945.7 49.4 51.2 45.4 46.4 46.9 49.2 46.3 PtIP-50Bb — — 76.5 85.9 47.448.9 47.4 49.6 52.9 46.9 48.4 49.7 52.2 47.6 PtIP-50Bc — — — 78.1 45.047.1 45.9 47.9 48.8 45.2 45.8 45.3 48.6 45.2 PtIP-50Bd — — — — 47.3 49.348.2 49.7 52.6 47.4 49.2 48.6 51.8 46.3 PtIP-50Fa — — — — — 44.0 39.542.6 58.4 40.9 40.2 49.9 55.6 40.9 PtIP-50Fb — — — — — — 52.3 71.7 46.951.2 51.4 44.5 45.4 77.9 PtIP-50Fd — — — — — — — 52.5 44.8 67.1 64.242.8 43.8 50.7 PtIP-50Fe — — — — — — — — 46.6 52.4 52.1 44.6 44.6 69.7PtIP-50Ff — — — — — — — — — 45.0 44.0 56.6 64.1 43.8 PtIP-50Fg — — — — —— — — — — 65.9 42.8 43.5 50.0 PtIP-50Fh — — — — — — — — — — — 43.5 44.050.0 PtIP-50Fi — — — — — — — — — — — — 60.8 42.1 PtIP-50Fj — — — — — — —— — — — — — 42.4 PtIP-50Fk — — — — — — — — — — — — — — PtIP-50Fl — — — —— — — — — — — — — — PtIP-50Fm — — — — — — — — — — — — — — PtIP-50Fn — —— — — — — — — — — — — — PtIP-50Fo — — — — — — — — — — — — — — PtIP-50Fp— — — — — — — — — — — — — — PtIP-50Fq — — — — — — — — — — — — — —PtIP-50Fr — — — — — — — — — — — — — — PtIP-50Fs — — — — — — — — — — — —— — PtIP-50Ft — — — — — — — — — — — — — — PtIP-50Ga — — — — — — — — — —— — — — PtIP-50Gb — — — — — — — — — — — — — — PtIP-50Gc — — — — — — — —— — — — — — PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP-PtIP- PtIP- PtIP- 50Fl 50Fm 50Fn 50Fo 50Fp 50Fq 50Fr 50Fs 50Ft 50Ga 50Gb50Gc 50Gd PtIP-50Aa 47.6 46.6 48.1 45.1 41.4 49.4 46.2 47.7 51.1 32.933.4 35.2 33.5 PtIP-50Ba 47.8 46.4 47.8 44.8 40.7 49.1 45.7 47.8 50.531.5 32.1 33.4 32.3 PtIP-50Bb 48.0 47.8 49.1 45.4 41.7 50.7 47.1 47.852.3 32.4 33.3 33.8 33.7 PtIP-50Bc 46.0 45.9 47.1 44.0 40.5 47.4 44.846.1 49.6 32.2 32.8 33.3 32.4 PtIP-50Bd 48.4 47.7 49.1 45.6 41.2 51.347.4 48.1 52.9 33.3 34.2 34.8 33.6 PtIP-50Fa 42.2 40.3 43.7 40.7 37.557.4 38.9 43.8 45.4 33.2 32.6 33.8 31.5 PtIP-50Fb 68.4 52.6 71.9 76.040.6 46.3 51.0 88.8 59.4 33.1 31.7 32.4 33.2 PtIP-50Fd 51.9 83.5 54.049.2 38.1 43.4 84.3 52.5 56.6 32.4 31.8 31.6 31.0 PtIP-50Fe 81.8 52.885.2 66.5 42.2 46.1 51.7 71.4 60.6 33.0 32.4 32.5 32.8 PtIP-50Ff 45.445.3 47.5 44.2 39.4 89.8 44.1 46.2 49.7 32.1 32.7 32.8 31.6 PtIP-50Fg50.8 66.0 53.3 48.6 38.4 44.3 65.7 51.4 56.7 31.0 29.6 32.3 30.4PtIP-50Fh 51.7 65.3 53.3 48.6 39.1 43.4 63.9 51.0 55.3 31.0 30.5 30.831.0 PtIP-50Fi 42.6 42.9 44.3 41.0 40.2 55.5 42.7 43.8 46.7 32.6 32.733.3 30.8 PtIP-50Fj 44.4 43.2 45.5 41.9 38.7 62.5 42.6 44.1 49.2 31.131.7 31.3 30.7 PtIP-50Fk 67.5 51.7 70.7 76.4 38.3 43.4 50.3 76.5 55.433.5 32.2 32.8 32.8 PtIP-50Fl — 52.0 86.2 63.3 40.9 45.2 51.1 68.0 57.832.3 31.3 32.4 32.5 PtIP-50Fm — — 54.3 51.2 39.4 44.6 84.7 53.3 56.532.6 31.8 31.8 31.8 PtIP-50Fn — — — 66.2 41.2 45.9 52.7 71.2 60.9 33.332.5 32.5 33.4 PtIP-50Fo — — — — 37.9 44.2 48.7 79.4 54.1 32.5 32.6 32.732.7 PtIP-50Fp — — — — — 39.8 38.7 40.4 41.9 39.3 39.6 39.9 38.3PtIP-50Fq — — — — — — 44.2 45.7 48.2 31.3 32.4 32.2 30.6 PtIP-50Fr — — —— — — — 51.8 55.9 31.7 31.1 31.4 30.6 PtIP-50Fs — — — — — — — — 58.633.4 31.9 32.9 33.0 PtIP-50Ft — — — — — — — — — 33.5 32.6 34.0 32.9PtIP-50Ga — — — — — — — — — — 85.9 87.4 77.2 PtIP-50Gb — — — — — — — — —— — 90.4 75.2 PtIP-50Gc — — — — — — — — — — — — 77.6

Moreover, tables summarizing the global identity are presented in Table10 and Table 11. Percent identity values were calculated using ClustalWalgorithm in the ALIGNX® module of the Vector NTI® Program Suite(Invitrogen™ Corporation, Carlsbad, Calif.) with all default parameters.

TABLE 11 PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP- PtIP-PtIP- PtIP- 65Ba 65Bb 65Ca 65Fa 65Fb 65Ga 65Gb 65Gc 65Gd 65Ge 65Ha 65HbPtIP-65Aa 82.9 83.2 76.8 46.4 46.9 35.2 35.8 33.6 34.5 34.5 33.6 26.6PtIP-65Ba — 85.0 74.0 44.8 44.8 33.9 34.1 33.2 32.9 32.9 33.2 26.8PtIP-65Bb — — 73.1 44.2 45.4 33.0 33.2 32.3 32.1 32.1 32.3 28.4PtIP-65Ca — — — 46.1 46.6 33.8 34.8 33.2 33.6 33 3 32.7 28.0 PtIP-65Fa —— — — 96.8 41.7 42.3 42.8 44.0 43.8 43.4 28.1 PtIP-65Fb — — — — — 42.342.8 42.8 44.0 43.8 43.4 28.7 PtIP-65Ga — — — — — — 98.6 85.1 83.3 83.183.9 27.2 PtIP-65Gb — — — — — — — 85.6 83.9 83.6 84.5 27.4 PtIP-65Gc — —— — — — — — 92.1 91.8 98.3 27.1 PtIP-65Gd — — — — — — — — — 99.7 92.725.9 PtIP-65Ge — — — — — — — — — — 92.4 25.7 PtIP-65Ha — — — — — — — — —— — 26.4 PtIP-65Hb — — — — — — — — — — — — PtIP-65Hc — — — — — — — — — —— — PtIP-65Hd — — — — — — — — — — — — PtIP-65He — — — — — — — — — — — —PtIP-65Hf — — — — — — — — — — — — PtIP-65Hg — — — — — — — — — — — —PtIP-65Hh — — — — — — — — — — — — PtIP-65Hj — — — — — — — — — — — —PtIP-65Hk — — — — — — — — — — — — PtIP- PtIP- PtIP- PtIP- PtIP- PtIP-PtIP- PtIP- 65Hc 65Hd 65He 65Hf 65Hg 65Hh 65Hj 65Hk PtIP-65Aa 28.5 28.126.8 27.5 27.5 29.4 30.7 30.2 PtIP-65Ba 29.2 28.5 27.8 28.0 28.6 28.529.3 30.1 PtIP-65Bb 31.8 29.4 27.7 29.7 28.6 27.1 29.0 30.4 PtIP-65Ca30.1 28.6 27.5 29.3 30.0 30.6 30.9 30.2 PtIP-65Fa 30.3 28.4 31.6 32.232.9 36.6 38.2 34.1 PtIP-65Fb 31.0 28.4 31.7 31.0 32.6 36.8 38.5 33.8PtIP-65Ga 23.2 24.4 23.4 24.6 29.4 31.6 56.0 32.2 PtIP-65Gb 23.8 24.923.6 24.9 29.4 31.3 56.3 31.7 PtIP-65Gc 23.8 23.6 24.1 25.1 29.7 31.253.5 30.8 PtIP-65Gd 23.3 23.4 24.7 25.9 29.0 32.5 56.2 31.7 PtIP-65Ge23.1 23.2 24.4 25.7 29.0 32.5 56.0 31.4 PtIP-65Ha 23.2 23.1 24.4 25.429.2 31.4 54.0 30.5 PtIP-65Hb 68.5 68.7 51.4 52.2 22.0 24.0 24.2 24.5PtIP-65Hc — 84.4 53.2 53.8 21.5 22.9 21.3 22.5 PtIP-65Hd — — 51.4 52.622.3 22.9 22.0 24.2 PtIP-65He — — — 81.7 23.4 23.7 23.6 24.0 PtIP-65Hf —— — — 24.0 22.1 23.8 26.1 PtIP-65Hg — — — — — 40.1 27.0 29.9 PtIP-65Hh —— — — — — 29.1 28.3 PtIP-65Hj — — — — — — — 29.3 PtIP-65Hk — — — — — — ——

Example 4: Transient Expression and Insect Bioassay on Transient LeafTissues

To confirm activity of PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ IDNO: 22) as working together as a complex, both were cloned into atransient expression system under control of a viral promoter dMMV andAtUBQ10 respectively (Day, et. al., (1999) Plant Mol. Biol. 40:771-782;Norris S R et al (1993) Plant Mol Biol. 21(5):895-906). PtIP-50Aa (SEQID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) were infiltrated alone andco-infiltrated at a 1:1 ratio. The polynucleotide encoding PtIP-65Aa(SEQ ID NO: 1) was cloned as a C-terminal HA tagged fusion vector fordetection purposes resulting in the polynucleotide of SEQ ID NO: 116encoding the polypeptide PtIP-65Aa-2×HA tag (SEQ ID NO: 217). Theagro-infiltration method of introducing an Agrobacterium cell suspensionto plant cells of intact tissues so that reproducible infection andsubsequent plant derived transgene expression may be measured or studiedis well known in the art (Kapila, et. al., (1997) Plant Science122:101-108). Briefly, the unifoliate stage of bush bean (common bean,Phaseolus vulgaris) or soybean (Glycine max), were agro-infiltrated withnormalized bacterial cell cultures of test and control strains. Leafdiscs were excised from each plantlet and infested with 2 neonates ofSoy Bean Looper (SBL) (Pseudoplusia includens), Corn Earworm, (CEW)(Helicoverpa zea) or 4 neonates of European Corn Borer (ECB) (Ostrinianubialis) alone. Leaf discs from a control was generated withAgrobacterium containing only a DsRed2 fluorescence marker (Clontech™,1290 Terra Bella Ave. Mountain View, Calif. 94043) expression vector.Leaf discs from a non-infiltrated plant was run as a second control. Theconsumption of green leaf tissue were scored after two (CEW) or three(ECB, SBL) days after infestation and given scores of 0 to 9. Neitherthe PtIP-50Aa polypeptide (SEQ ID NO: 71) or the PtIP-65Aa-2×HA taggedpolypeptide (SEQ ID NO: 217) alone showed insecticidal activity at theconcentrations tested, however insecticidal activity was observed whenthe expression vectors for both proteins were co-infiltrated (Table 12).The transiently co-expressed PtIP-50Aa polypeptide (SEQ ID NO: 71) andPtIP-65Aa-2×HA tag polypeptide (SEQ ID NO: 217) protected leaf discsfrom consumption by the infested insects while total green tissueconsumption was observed for the negative control and untreated tissue.Transient protein expression of both PtIP-50Aa (SEQ ID NO: 71) andPtIP-65Aa-2×HA tagged (SEQ ID NO: 217) was confirmed by massspectrometry-based protein identification method using trypsinizedprotein extracts of infiltrated leaf tissues (Patterson, (1998)10(22):1-24, Current Protocol in Molecular Biology published by JohnWiley & Son Inc) and by Western Analysis of the HA tag. These proteinswere also active when run as a molecular stack in the bush beantransient assay system.

Experiments were run to test various tagged versions of both PtIP-50Aa(PtIP50Aa-GAAEPEA tag SEQ ID NO: 207; MBP-rTEV-PtIP50Aa SEQ ID NO: 219)and PtIP-65Aa (PtIP065Aa-GAAEPEA tag SEQ ID NO: 213; PtIP65Aa-rTEV-2×HAtag SEQ ID NO: 215; 2×HA tag-PtIP65Aa SEQ ID NO: 209; 10HIS-MBP-rTEV-PtIP65Aa SEQ ID NO: 221) which showed that tagged versionsof both proteins are still active and similar to untagged protein.

TABLE 12 Construct Insect Avg. StdDEV PtIP-50Aa (SEQ ID NO: 71) + SBL 90 PtIP-65Aa-2XHA tag (SEQ ID NO: 217) (1:1) pVER SBL 2.8 2.3 blank SBL 10 PtIP-50Aa (SEQ ID NO: 71) + CEW 9 0 PtIP-65Aa-2XHA tag (SEQ ID NO:217) (1:1) pVER CEW 1 0 blank CEW 1 0 PtIP-50Aa (SEQ ID NO: 71) + ECB5.8 1 PtIP-65Aa-2XHA tag (SEQ ID NO: 217) (1:1) pVER ECB 1.4 1 blank ECB1.4 1 Value Description 1 leaf disk is greater than 90% consumed 2 leafdisk is 70-80% consumed 3 leaf disk is 60-70% consumed 4 leaf disk is50-60% consumed 5 leaf disk is 40-50% consumed 6 leaf disk is less than30% consumed 7 leaf disk is less than 10% consumed 8 leaf disk has onlya few pinholes 9 leaf disk is untouched by the insect

Example 5: Lepidoptera Assays with Purified Proteins TransientlyExpressed in Bush Bean Assay

Insecticidal activity bioassay screens were conducted with C-terminallyHA tagged PtIP-65Aa-rTEV-2×HA (SEQ ID NO: 215), and PtIP-50Aa (SEQ IDNO: 71) that were co-infiltrated into the bush bean transient systemdescribed above. The protein was purified from homogenized plant tissueusing affinity beads targeting the 2×HA tag (Sigma-Aldrich A2095Monoclonal Anti-HA-Agarose). The protein was tested in diet assay toevaluate the insecticidal protein effects on larvae of a diversity ofLepidoptera.

Lepidoptera feeding assays were conducted on an artificial dietcontaining PtIP-50Aa protein (SEQ ID NO: 71) and PtIP-65Aa-rTEV-2×HA(SEQ ID NO: 215) from 1:1 infiltrated Bush Bean. The PtIP-50Aa (SEQ IDNO: 71) and PtIP-65Aa-rTEV-2×HA (SEQ ID NO: 215) insecticidal proteinswere tested for activity against Soy Bean Looper (SBL) (Pseudoplusiaincludens), Corn Earworm, (CEW) (Helicoverpa zea) and European CornBorer (ECB) (Ostrinia nubialis). For SBL, the proteins were incorporatedinto a Lepidopteran-specific artificial diet (Southland Products) wellsat a rate of 0, 0.06 ppm, 0.2 ppm, 0.63 ppm, 2 ppm, 6.3 ppm and 20 ppm.For CEW, the proteins were incorporated into a Lepidopteran-specificartificial diet (Southland Products) wells at a rate of 0, 0.16 ppm, 0.5ppm, 1.58 ppm, 5.0 ppm, 15.8 ppm and 50 ppm. For ECB, the proteins wereincorporated into a Lepidopteran-specific artificial diet (SouthlandProducts) wells at a rate of 0, 0.38 ppm, 1.2 ppm, 3.8 ppm, 12.0 ppm, 38ppm and 120 ppm. Neonate (2-3) larvae were placed in each well to feedfor four days. Each bioassay was run with 4 duplicates at each dose.Results were expressed as positive for larvae reactions such as stuntingand or mortality. Results were expressed as negative if the larvae weresimilar to the negative control that was fed diet to which the abovebuffer only has been applied. Larvae that had a size similar to controlswere assigned a score of 0, those that showed slight stunting werescored 1, those that showed severe stunting were scored 2 and those thatwere dead were scored 3. The insecticidal activity of PtIP-50Aa (SEQ IDNO: 71) and PtIP-65Aa-rTEV-2×HA (SEQ ID NO: 215) combined in the SoyBean Looper (SBL) (Pseudoplusia includens), Corn Earworm, (CEW)(Helicoverpa zea) and European Corn Borer (ECB) (Ostrinia nubialis) isshown in Table 13. Table 13 shows the LC50 based on the concentration(ppm) of PtIP-50Aa (SEQ ID NO: 71).

TABLE 13 PtIP-50Aa (SEQ ID NO: 71) Lower Upper Insect ppm 95% CL 95% CLCEW LC50 16.73 12.50 23.18 ECB LC50 >120 SBL LC50 0.9196 0.3099 2.578

Example 6: Co-Infiltration of PtIP-50 Polypeptides and PtIP-65Polypeptides Cloned from Different Genera of Ferns Exhibit Activity

Polynucleotides encoding PtIP-50 polypeptide homologs and PtIP-65polypeptide homologs were identified from several species of ferns asshown in Example 3 and some were cloned into the transient expressionvector as described in Example 2 above. Polynucleotides encoding thehomologs were cloned from one of three ferns, Asplenium australasicum,Polypodium musifolium and Adiantum pedatum (Table 14.) Each homolog wasco-infiltrated 1:1 in the bush bean transient system in a mix and matchexperiment to determine activity and spectrum from each combination.Results from various combinations in the mix and match experiment showthat PtIP-50 polypeptides and PtIP-65 polypeptides from differentsources of ferns can have activity with each other (Table 15.) Thespectrum of activity is broadened with the improvement in FAW activitywith PtIP-65Ha (SEQ ID NO: 25).

TABLE 14 Fern PtIP-50 Homolog PtIP-65 Homolog Asplenium australasicumPtIP-50Aa PtIP-65Aa (SEQ ID NO: 43) (SEQ ID NO: 1) Polypodium musifoliumPtIP-50Ba PtIP-65Ba (SEQ ID NO: 49) (SEQ ID NO: 3) PtIP-65Ha (SEQ ID NO:4) Adiantum pedatum PtIP-50Bb PtIP-65Ca (SEQ ID NO: 56) (SEQ ID NO: 8)

TABLE 15 PtIP-50 PtIP-65 bushbean transient activity polypeptidepolypeptide CEW ECB FAW SBL PtIP-50Aa PtIP-65Aa 8.3 6.3 1.6 8.9 (SEQ IDNO: 71) (SEQ ID NO: 22) PtIP-50Aa PtIP-65Ba 8.5 1.3 1.8 6.9 (SEQ ID NO:71) (SEQ ID NO: 24) PtIP-50Aa PtIP-65Ca 8.0 8.4 1.6 9.0 (SEQ ID NO: 71)(SEQ ID NO: 29) PtIP-50Aa PtIP-65Ha 5.8 1.2 1.0 8.5 (SEQ ID NO: 71) (SEQID NO: 25) PtIP-50Ba PtIP-65Ba 8.8 1.6 1.0 6.3 (SEQ ID NO: 77) (SEQ IDNO: 24) PtIP-50Bb PtIP-65Aa 8.4 6.4 1.0 9.0 (SEQ ID NO: 84) (SEQ ID NO:22) PtIP-50Bb PtIP-65Ba 8.3 2.2 1.0 5.3 (SEQ ID NO: 84) (SEQ ID NO: 24)PtIP-50Bb PtIP-65Ca 8.4 9.0 1.0 9.0 (SEQ ID NO: 84) (SEQ ID NO: 29)PtIP-50Ga PtIP-65Hb 6.8 7.8 1.8 8.0 (SEQ ID NO: 88) (SEQ ID NO: 31)

Example 7: Baculovirus Expression of PtIP-50Aa and PtIP-65AaPolypeptides

The gene encoding PtIP-50Aa (SEQ ID NO: 71) was synthesized andsubcloned into a pFastBac™ Dual vector (Invitrogen®) with the PtIP-50Aanative stop codon (TAA) (SEQ ID NO: 100). The gene encodingPtIP-65Aa-10×His (SEQ ID NO: 223) was subcloned with the stop codonremoved for C-terminal translation of a 10×-histidine tag addition andthe gene sequence of the histidine-tagged PtIP-65Aa is set forth as SEQID NO: 122. This vector was transformed into DH10Bac cells to generatebaculovirus. These baculovirus were used to infect sf9 insect cells andincubated for 72 hours at 27° C. The infected insect cells wereharvested by centrifugation. The cell culture pellet was suspended with200 ml of lysis buffer (1×PBS, 10% glycerol, with protease inhibitor andbenzonase) and incubated at 4° C. for 5 min with stirring, thenhomogenizing twice. Chaps was added to lysate to a 1.0% finalconcentration and this was stirred at 4° C. for 1 hour. The lysate wascentrifuged at 16000 rpm for 20 min. The supernatant was saved andloaded onto six 2 ml Ni-NTA columns pre-equilibrated with Elute buffer(1×PBS, 10% glycerol). The columns were then eluted with 10 ml of Elutebuffer consisting of 10, 20, 50, and 250 mM of imidazole. Samples wereanalyzed by SDS-PAGE. The fraction from the 250 mM imidazole elutionwere concentrated to 5 mL and loaded onto two PD-10 desalting columns(GE Life Sciences, Pittsburgh, USA) pre-equilibrated with Elute buffer.3 mL of Elute buffer was loaded on to each column and 2.5 mL of eluatecollected from each column. The eluates from two desalting columns werecombined. Lysates were assayed against SBL and CEW and were active whenboth PtIP-65Aa-10×His (SEQ ID NO: 223) and PtIP-50Aa (SEQ ID NO: 71)were present (Table 16).

TABLE 16 Protein & LC/IC50 Lower Upper expression Insect LC/IC ppm (4 d)95% CL 95% CL PtIP-50Aa CEW LC50 >100 (SEQ ID NO: 71)/ IC50 22.24 12.5647.10 65Aa-10XHis SBL LC50 1.157 0.7220 1.811 (SEQ ID NO: 223) IC500.2992 0.1974 0.4297

Example 8: Agrobacterium-Mediated Transformation of Maize andRegeneration of Transgenic Plants

For Agrobacterium-mediated transformation of maize with PtIP-65 andPtIP-50 nucleotide sequences such as PtIP-50Aa (SEQ ID NO: 43) andPtIP-65Aa (SEQ ID NO: 1), the method of Zhao can be used (U.S. Pat. No.5,981,840 and PCT Patent Publication Number WO 1998/32326; the contentsof which are hereby incorporated by reference). Briefly, immatureembryos are isolated from maize and the embryos contacted with asuspension of Agrobacterium under conditions whereby the bacteria arecapable of transferring the nucleotide sequence (PtIP-50Aa SEQ ID NO:43, PtIP-65Aa SEQ ID NO: 1) to at least one cell of at least one of theimmature embryos (step 1: the infection step). In this step the immatureembryos can be immersed in an Agrobacterium suspension for theinitiation of inoculation. The embryos are co-cultured for a time withthe Agrobacterium (step 2: the co-cultivation step). The immatureembryos can be cultured on solid medium following the infection step.Following this co-cultivation period an optional “resting” step iscontemplated. In this resting step, the embryos are incubated in thepresence of at least one antibiotic known to inhibit the growth ofAgrobacterium without the addition of a selective agent for planttransformation (step 3: resting step). The immature embryos can becultured on solid medium with antibiotic, but without a selecting agent,for elimination of Agrobacterium and for a resting phase for theinfected cells. Next, inoculated embryos are cultured on mediumcontaining a selective agent and growing transformed callus is recovered(step 4: the selection step). The immature embryos are cultured on solidmedium with a selective agent resulting in the selective growth oftransformed cells. The callus is then regenerated into plants (step 5:the regeneration step), and calli grown on selective medium can becultured on solid medium to regenerate the plants.

Example 9: Transformation and Regeneration of Soybean (Glycine max)

Transgenic soybean lines are generated by the method of particle gunbombardment (Klein et al., Nature (London) 327:70-73 (1987); U.S. Pat.No. 4,945,050) using a BIORAD Biolistic PDS1000/He instrument and eitherplasmid or fragment DNA. The following stock solutions and media areused for transformation and regeneration of soybean plants:

Stock Solutions:

Sulfate 100× Stock:

-   -   37.0 g MgSO₄.7H₂O, 1.69 g MnSO₄.H₂O, 0.86 g ZnSO₄.7H₂O, 0.0025 g        CuSO₄.5H₂O        Halides 100× Stock:    -   30.0 g CaCl₂.2H₂O, 0.083 g KI, 0.0025 g CoCl₂.6H₂O        P, B, Mo 100× Stock:    -   18.5 g KH₂PO₄, 0.62 g H₃BO₃, 0.025 g Na₂MoO₄.2H₂O        Fe EDTA 100× Stock:    -   3.724 g Na₂EDTA, 2.784 g FeSO₄.7H₂O        2,4-D Stock:    -   10 mg/mL Vitamin        B5 vitamins, 1000× Stock:        100.0 g myo-inositol, 1.0 g nicotinic acid, 1.0 g pyridoxine        HCl, 10 g thiamine.HCL.        Media (Per Liter):        S13199 Solid Medium:        1 package MS salts (Gibco/BRL Cat. No. 11117-066), 1 mL B5        vitamins 1000× stock, 30 g Sucrose, 4 ml 2, 4-D (40 mg/L final        concentration), pH 7.0, 2 gm Gelrite        SB1 Solid Medium:        1 package MS salts (Gibco/BRL Cat. No. 11117-066), 1 mL B5        vitamins 1000× stock, 31.5 g Glucose, 2 mL 2, 4-D (20 mg/L final        concentration), pH 5.7, 8 g TC agar        SB196:        10 mL of each of the above stock solutions 1-4, 1 mL B5 Vitamin        stock, 0.463 g (NH4)2 SO4, 2.83 g KNO3, 1 mL 2, 4 D stock, 1 g        asparagine, 10 g Sucrose, pH 5.7        SB71-4:        Gamborg's B5 salts, 20 g sucrose, 5 g TC agar, pH 5.7.        S13103:        1 pk. Murashige & Skoog salts mixture, 1 mL B5 Vitamin stock,        750 mg MgCl2 hexahydrate, 60 g maltose, 2 g gelrite, pH 5.7.        SB166:        SB103 supplemented with 5 g per liter activated charcoal.        Soybean Embryogenic Suspension Culture Initiation:

Pods with immature seeds from available soybean plants 45-55 days afterplanting are picked, removed from their shells and placed into asterilized magenta box. The soybean seeds are sterilized by shaking themfor 15 min in a 5% Clorox® solution with 1 drop of Ivory™ soap (i.e., 95mL of autoclaved distilled water plus 5 mL Clorox® and 1 drop of soap,mixed well). Seeds are rinsed using 2 L sterile distilled water andthose less than 3 mm are placed on individual microscope slides. Thesmall end of the seed is cut and the cotyledons pressed out of the seedcoat. Cotyledons are transferred to plates containing SB199 medium(25-30 cotyledons per plate) for 2 weeks, then transferred to SB1 for2-4 weeks. Plates are wrapped with fiber tape. After this time,secondary embryos are cut and placed into SB196 liquid medium for 7days.

Culture Conditions:

Soybean embryogenic suspension cultures (cv. 93Y21) were maintained in50 mL liquid medium SB196 on a rotary shaker, 100-150 rpm, 26° C. on16:8 h day/night photoperiod at light intensity of 80-100 pE/m2/s.Cultures are subcultured every 7-14 days by inoculating up to ½ dimesize quantity of tissue (clumps bulked together) into 50 mL of freshliquid SB196.

Preparation of DNA for Bombardment:

In particle gun bombardment procedures it is possible to use purified 1)entire plasmid DNA; or 2) DNA fragments containing only the recombinantDNA expression cassette(s) of interest. For every seventeen bombardmenttransformations, 85 μL of suspension is prepared containing 1 to 90picograms (pg) of plasmid DNA per base pair of each DNA plasmid. DNAplasmids or fragments are co-precipitated onto gold particles asfollows. The DNAs in suspension are added to 50 μL of a 10-60 mg/mL 0.6μm gold particle suspension and then combined with 50 μL CaCl₂ (2.5 M)and 20 μL spermidine (0.1 M). The mixture is vortexed for 5 sec, spun ina microfuge for 5 sec, and the supernatant removed. The DNA-coatedparticles are then washed once with 150 μL of 100% ethanol, vortexed andspun in a microfuge again, then resuspended in 85 μL of anhydrousethanol. Five μL of the DNA-coated gold particles are then loaded oneach macrocarrier disk.

Tissue Preparation and Bombardment with DNA:

Approximately 100 mg of two-week-old suspension culture is placed in anempty 60 mm×15 mm petri plate and the residual liquid removed from thetissue using a pipette. The tissue is placed about 3.5 inches away fromthe retaining screen and each plate of tissue is bombarded once.Membrane rupture pressure is set at 650 psi and the chamber is evacuatedto 28 inches of Hg. Following bombardment, the tissue from each plate isdivided between two flasks, placed back into liquid media, and culturedas described above.

Selection of Transformed Embryos and Plant Regeneration:

After bombardment, tissue from each bombarded plate is divided andplaced into two flasks of SB196 liquid culture maintenance medium perplate of bombarded tissue. Seven days post bombardment, the liquidmedium in each flask is replaced with fresh SB196 culture maintenancemedium supplemented with 100 ng/ml selective agent (selection medium).For selection of transformed soybean cells the selective agent used canbe a sulfonylurea (SU) compound with the chemical name,2-chloro-N-((4-methoxy-6 methy-1,3,5-triazine-2-yl)aminocarbonyl)benzenesulfonamide (common names: DPX-W4189 and chlorsulfuron).Chlorsulfuron is the active ingredient in the DuPont sulfonylureaherbicide, GLEAN®. The selection medium containing SU is replaced everytwo weeks for 8 weeks. After the 8 week selection period, islands ofgreen, transformed tissue are observed growing from untransformed,necrotic embryogenic clusters. These putative transgenic events areisolated and kept in SB196 liquid medium with SU at 100 ng/ml foranother 5 weeks with media changes every 1-2 weeks to generate new,clonally propagated, transformed embryogenic suspension cultures.Embryos spend a total of around 13 weeks in contact with SU. Suspensioncultures are subcultured and maintained as clusters of immature embryosand also regenerated into whole plants by maturation and germination ofindividual somatic embryos.

Somatic embryos became suitable for germination after four weeks onmaturation medium (1 week on SB166 followed by 3 weeks on SB103). Theyare then removed from the maturation medium and dried in empty petridishes for up to seven days. The dried embryos are then planted inSB71-4 medium where they are allowed to germinate under the same lightand temperature conditions as described above. Germinated embryos aretransferred to potting medium and grown to maturity for seed production.

Leaf disc were excised from the transformed plants and tested forinsecticidal activity of PtIP-50 and PtIP-65 against the Soy Bean Looper(SBL) (Pseudoplusia includens), Corn Earworm, (CEW) (Helicoverpa zea),European Corn Borer (ECB) (Ostrinia nubialis) and Velvet BeanCaterpillar (VBC) (Anticarsia gemmatalis). The expected activity wasobserved for SBL, CEW and ECB, however activity was recorded in assayagainst VBC as well (Table 17).

TABLE 17 VBC 3rd SBL 3rd SBL NEO CEW NEO FAW NEO VBC 1st ECB NEO EventDNA avg. score avg. score avg. score avg. score avg. score avg. scoreavg. score SOY 1985.3.1 PHP61857A 5.5 8.0 8.3 8.0 1.0 8.0 8.7 SOY1987.2.1 PHP61857A 4.5 8.0 8.8 8.3 1.0

Example 10: Mode of Action

PtIP-50 polypeptides and PtIP-65 polypeptides were evaluated forstability in the presence of midgut fluid extracts from Pseudoplusiaincludens to determine if their full length states represent pro-formsof the proteins and whether midgut proteolysis is required foractivation to a toxic state in vivo.

The ability of the mixture of a PtIP-50 polypeptide and a PtIP-65polypeptide to alter short circuit current (SCC) in isolated midgutsfrom the Lepidoptera, Anacarsia gemmatalis was evaluated. Effects on theSCC in this system is considered consistent with pore formation, ionchannel interaction, electrogenic transporter interaction, or possiblydisruption of tight junctions that maintain diffusional barriers betweenmidgut cells. An example of the effects of the mixture of the PtIP-50Aa(SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) polypeptides on SCC isshown in FIG. 18. The ISCC reflects the transport activity and midgutstructural integrity to maintain normal midgut function. The decline inISCC following the addition of PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa(SEQ ID NO: 22) polypeptides reflects the loss of ionic balance in themidgut.

The direct binding of the PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQID NO: 22) polypeptides to Pseudoplusia includens brush border membranevesicles was tested for target site identification. Specific binding,demonstrates the presence of a receptor for this binary. Heterologouscompetition with other insect-active toxins was tested to evaluate thecompetitive binding to the target site. Specific binding of PtIP-50Aa(SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) is illustrated in FIGS. 19and 20 a and 20 b. Specific binding, demonstrates the presence of areceptor for this binary. Heterologous competition with otherinsect-active toxins was tested to evaluate the competitive binding tothe target site. Specific binding of PtIP-50Aa (SEQ ID NO: 71) andPtIP-65Aa (SEQ ID NO: 22) is illustrated in FIGS. 19, 20 a and 20 b.FIG. 19 shows a gel image of the specific binding of PtIP-50Aa (SEQ IDNO: 71) and PtIP-65Aa (SEQ ID NO: 22) to BBMVs from C. includens. As theconcentration of unlabeled PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQID NO: 22) is increased, the likelihood that Alexa-labeled PtIP-50Aa(SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) can bind to the targetreceptor is decreased. The concentration at which the fluorescencesignal from bound Alexa-PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ IDNO: 22) is reduced to 50% when compared to the signal measured in theabsence of unlabeled PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO:22) reflects the point at which half (50%) of the total receptors areoccupied by unlabeled PtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ IDNO: 22) and is referred to as the EC50 concentration. Densitometryresults and EC50 determinations from the average three replicates areshown in FIGS. 20a and 20b . FIGS. 20a and 20b show the evaluation ofEC50 values from densitometry of gel images for PtIP-50Aa (SEQ ID NO:71) and PtIP-65Aa (SEQ ID NO: 22) binding to C. includens BBMVs. FIG.20a shows the average densitometry values for bound Alexa-PtIP-50Aa (SEQID NO: 71) in the presence of different concentrations of unlabeledPtIP-50Aa (SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) normalized tothe amount bound in the absence of unlabeled PtIP-50Aa (SEQ ID NO: 71)and PtIP-65Aa (SEQ ID NO: 22). The solid line reflects the best fit of asquare logistic equation to the data. The data are best fit by atwo-component equation having EC50 values of 10 nM and 370 nM for thehigh affinity and low affinity components, respectively. FIG. 20b showsthe average densitometry values for bound Alexa-PtIP-65Aa (SEQ ID NO:22) in the presence of different concentrations of unlabeled PtIP-50Aa(SEQ ID NO: 71) and PtIP-65Aa (SEQ ID NO: 22) normalized to the amountbound in the absence of unlabeled PtIP-50Aa (SEQ ID NO: 71) andPtIP-65Aa (SEQ ID NO: 22). The EC50 value from the fit is 14 nM.

Example 11: PtIP-50 & PtIP-65 Fusion Proteins

Fusion protein(s) of PtIP-50Aa and PtIP-65Aa (SEQ ID NO: 71 and SEQ IDNO: 22) Constructs on Excel Sheet) were created with a small syntheticlinker between the two proteins and cloned into the Bush Bean transientsystem.

Two fusion proteins were made with PtIP-50Aa either N-terminal orC-terminal to PtIP-65Aa. After infiltration of bush bean leaf discs wereinfested with either CEW, ECB or SBL and assayed for 3 (CEW) or 4 (SBL,ECB) days. The results of the assay show that the fusion protein didprotect the leaf discs from consumption and are a viable control option(Table 18).

TABLE 18 Fusion PtIP-65Aa/PtIP-50Aa SBL (9.0) CEW (7.2)PtIP-50Aa/PtIP-65Aa SBL (8.8) CEW (8.7) ECB (6.1)

Example 12—Particle Bombardment Transformation and Regeneration ofTransgenic Plants

Immature maize embryos from greenhouse donor plants are bombarded with aplasmid containing a nucleotide sequence encoding the insecticidalprotein. The ears are husked and surface sterilized in 30% Clorox®bleach plus 0.5% Micro detergent for 20 minutes and rinsed two timeswith sterile water. The immature embryos are excised and placed embryoaxis side down (scutellum side up), 25 embryos per plate, on 560Y mediumfor 4 hours and then aligned within the 2.5 cm target zone inpreparation for bombardment. A plasmid vector DNA comprising thenucleotide sequence encoding the insecticidal protein operably linked toa promoter is precipitated onto 1.1 μm (average diameter) tungstenpellets using a CaCl₂ precipitation procedure as follows: 100 μlprepared tungsten particles in water; 10 μl (1 μg) DNA in Tris EDTAbuffer (1 μg total DNA); 100 μl 2.5 M CaCl₂ and 10 μl 0.1 M spermidine.

Each reagent is added sequentially to the tungsten particle suspension,while maintained on the multitube vortexer. The final mixture issonicated briefly and allowed to incubate under constant vortexing for10 minutes. After the precipitation period, the tubes are centrifugedbriefly, liquid removed, washed with 500 ml 100% ethanol and centrifugedfor 30 seconds. Again the liquid is removed, and 105 μl 100% ethanol isadded to the final tungsten particle pellet. For particle gunbombardment, the tungsten/DNA particles are briefly sonicated and 10 μlspotted onto the center of each macrocarrier and allowed to dry about 2minutes before bombardment. The sample plates are bombarded at level #4in a particle gun. All samples receive a single shot at 650 PSI, with atotal of ten aliquots taken from each tube of prepared particles/DNA

Following bombardment, the embryos are kept on 560Y medium for 2 days,then transferred to 560R selection medium containing 3 mg/literBialaphos, and subcultured every 2 weeks. After approximately 10 weeksof selection, selection-resistant callus clones are transferred to 288Jmedium to initiate plant regeneration. Following somatic embryomaturation (2-4 weeks), well-developed somatic embryos are transferredto medium for germination and transferred to the lighted culture room.Approximately 7-10 days later, developing plantlets are transferred to272V hormone-free medium in tubes for 7-10 days until plantlets are wellestablished. Plants are then transferred to inserts in flats (equivalentto 2.5″ pot) containing potting soil and grown for 1 week in a growthchamber, subsequently grown an additional 1-2 weeks in the greenhouse,then transferred to classic 600 pots (1.6 gallon) and grown to maturity.Plants are monitored and scored for expression of a PtIP-50 polypeptideor a PtIP-65 polypeptide by assays known in the art, such as, forexample, immunoassays and Western blotting.

Transgenic maize plants positive for expression of the insecticidalproteins are tested for pesticidal activity using standard bioassaysknown in the art. Such methods include, for example, root excisionbioassays and whole plant bioassays. See, e.g., US Patent ApplicationPublication Number US 2003/0120054 and International Publication NumberWO 2003/018810.

Bombardment medium (560Y) comprises 4.0 g/l N6 basal salts (SIGMAC-1416), 1.0 ml/l Eriksson's Vitamin Mix (1000.times.SIGMA-1511), 0.5mg/l thiamine HCl, 120.0 g/l sucrose, 1.0 mg/l 2,4-D and 2.88 g/lL-proline (brought to volume with D-I H₂O following adjustment to pH 5.8with KOH); 2.0 g/l Gelrite (added after bringing to volume with D-I H₂O)and 8.5 mg/l silver nitrate (added after sterilizing the medium andcooling to room temperature). Selection medium (560R) comprises 4.0 g/lN6 basal salts (SIGMA C-1416), 1.0 ml/l Eriksson's Vitamin Mix(1000.times.SIGMA-1511), 0.5 mg/l thiamine HCl, 30.0 g/l sucrose and 2.0mg/l 2,4-D (brought to volume with D-I H₂O following adjustment to pH5.8 with KOH); 3.0 g/l Gelrite (added after bringing to volume with D-IH₂O) and 0.85 mg/l silver nitrate and 3.0 mg/l bialaphos (both addedafter sterilizing the medium and cooling to room temperature).

Plant regeneration medium (288J) comprises 4.3 g/l MS salts (GIBCO11117-074), 5.0 ml/l MS vitamins stock solution (0.100 g nicotinic acid,0.02 g/l thiamine HCL, 0.10 g/l pyridoxine HCL, and 0.40 g/l glycinebrought to volume with polished D-I H₂O) (Murashige and Skoog, (1962)Physiol. Plant. 15:473), 100 mg/l myo-inositol, 0.5 mg/l zeatin, 60 g/lsucrose and 1.0 ml/l of 0.1 mM abscisic acid (brought to volume withpolished D-I H₂O after adjusting to pH 5.6); 3.0 g/l Gelrite (addedafter bringing to volume with D-I H₂O) and 1.0 mg/l indoleacetic acidand 3.0 mg/l bialaphos (added after sterilizing the medium and coolingto 60° C.). Hormone-free medium (272V) comprises 4.3 g/l MS salts (GIBCO11117-074), 5.0 ml/l MS vitamins stock solution (0.100 g/l nicotinicacid, 0.02 g/l thiamine HCL, 0.10 g/l pyridoxine HCL and 0.40 g/lglycine brought to volume with polished D-I H₂O), 0.1 g/l myo-inositoland 40.0 g/l sucrose (brought to volume with polished D-I H₂O afteradjusting pH to 5.6) and 6 g/l bacto-agar (added after bringing tovolume with polished D-I H₂O), sterilized and cooled to 60° C.

The above description of various illustrated embodiments of thedisclosure is not intended to be exhaustive or to limit the scope to theprecise form disclosed. While specific embodiments of and examples aredescribed herein for illustrative purposes, various equivalentmodifications are possible within the scope of the disclosure, as thoseskilled in the relevant art will recognize. The teachings providedherein can be applied to other purposes, other than the examplesdescribed above. Numerous modifications and variations are possible inlight of the above teachings and, therefore, are within the scope of theappended claims.

These and other changes may be made in light of the above detaileddescription. In general, in the following claims, the terms used shouldnot be construed to limit the scope to the specific embodimentsdisclosed in the specification and the claims.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books or otherdisclosures) in the Background, Detailed Description, and Examples isherein incorporated by reference in their entireties.

Efforts have been made to ensure accuracy with respect to the numbersused (e.g. amounts, temperature, concentrations, etc.) but someexperimental errors and deviations should be allowed for. Unlessotherwise indicated, parts are parts by weight, molecular weight isaverage molecular weight; temperature is in degrees centigrade; andpressure is at or near atmospheric.

That which is claimed is:
 1. A recombinant PtIP-50 polypeptidecomprising an amino acid sequence having at least 95% sequence identity,across the entire length of the amino acid sequence, compared to theamino acid sequence of SEQ ID NO: 71, wherein the PtIP-50 polypeptidehas insecticidal activity against Soy Bean Looper, Corn Earworm,European Corn Borer or Velvet Bean Caterpillar in combination with aPtIP-65 polypeptide comprising an amino acid sequence having at least95% identity to SEQ ID NO: 22, wherein the PtIP-50 polypeptide isoperably linked to a heterologous signal or leader sequence.
 2. Therecombinant PtIP-50 polypeptide of claim 1, wherein the PtIP-50polypeptide further comprises one or more characteristic selected fromthe group consisting of: a) a molecular weight of between about 80 kDaand about 100 kDa; b) an isoelectric point between about 4.4 and 5.2;and c) does not have chitinase activity.
 3. A recombinant PtIP-50polypeptide comprising an amino acid sequence having at least 95%sequence identity, across the entire length of the amino acid sequence,compared to the amino acid sequence of SEQ ID NO: 71, wherein thePtIP-50 polypeptide has insecticidal activity against Soy Bean Looper,Corn Earworm, European Corn Borer or Velvet Bean Caterpillar incombination with a PtIP-65 polypeptide comprising an amino acid sequencehaving at least 95% identity to SEQ ID NO: 22, and the PtIP-50polypeptide, in combination with the PtIP-65 polypeptide, specificallybinds to brush border membrane vesicles isolated from Soy Bean Looper.4. The recombinant PtIP 50 polypeptide of claim 1, wherein A recombinantPtIP-50 polypeptide comprising an amino acid sequence having at least95% sequence identity, across the entire length of the amino acidsequence, compared to the amino acid sequence of SEQ ID NO: 71, whereinthe PtIP-50 polypeptide has insecticidal activity against Soy BeanLooper, Corn Earworm, European Corn Borer or Velvet Bean Caterpillar incombination with a PtIP-65 polypeptide comprising an amino acid sequencehaving at least 95% identity to SEQ ID NO: 22, and the PtIP-50polypeptide, in combination with the PtIP-65 polypeptide, disrupts theionic balance across brush border membrane vesicles isolated from SoyBean Looper.
 5. A polynucleotide encoding a PtIP-50 polypeptidecomprising an amino acid sequence having at least 95% sequence identity,across the entire length of the amino acid sequence, compared to theamino acid sequence of SEQ ID NO: 71, wherein the PtIP-50 polypeptidehas insecticidal activity against Soy Bean Looper, Corn Earworm,European Corn Borer or Velvet Bean Caterpillar in combination with aPtIP-65 polypeptide comprising an amino acid sequence having at least95% identity to SEQ ID NO: 22, wherein the polynucleotide encoding thePtIP-50 polypeptide is operably linked to a heterologous regulatoryelement.
 6. A fusion protein comprising: a PtIP-50 polypeptidecomprising: an amino acid sequence having at least 95% sequenceidentity, across the entire length of the amino acid sequence, comparedto the amino acid sequence of SEQ ID NO: 71, wherein the PtIP-50polypeptide has insecticidal activity against Soy Bean Looper, CornEarworm, European Corn Borer or Velvet Bean Caterpillar in combinationwith a PtIP-65 polypeptide comprising an amino acid sequence having atleast 95% identity to SEQ ID NO: 22; and a polypeptide selected from: a)a PtIP-65 polypeptide comprising an amino acid sequence having at least95% sequence identity, across the entire length of the amino acidsequence, compared to the amino acid sequence of SEQ ID NO: 22; b) aninsecticidal polypeptide having a different mode of action from thePtIP-50 polypeptide; and c) a polypeptide to increase expression,facilitate purification or detection.